Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
YY2 controls CD276 transcription in Tsc2-deficient cells. Enhanced Cd276 promoter activity in Tsc2−/− 105K cells expressing empty vector (EV) compared to reconstitution of TSC2 (a) and in Tsc2 KO MEFs compared to Tsc2-WT MEFs (b). Relative luciferase activity was determined by a dual-luciferase assay system. psiCHECK2-Cd276 encodes the Cd276 promoter. Empty psiCHECK2 vector was used as the negative control. n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Raptor, mTOR or S6K knockdown suppresses Cd276 promoter activity in Tsc2−/− 105K cells (c) and Tsc2 KO MEFs (d). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. siRNA knockdown of YY2 reduces B7-H3 protein expression in Tsc2−/− 105K cells (e) and Tsc2 KO MEFs (f) (n = 3). Knockdown of YY2 reduces Cd276 mRNA expression in Tsc2−/− 105K cells (g) and Tsc2 KO MEFs (h). n = 3, means ± SD, two-tailed unpaired Student’s t-test, *p < 0.05, ***p < 0.001. i Promoter region of Cd276 displaying the location of the YY2 binding site. YY2 ChIP-qPCR analysis showing increased YY2 occupancy on the Cd276 promoter in Tsc2−/− 105K cells (j) and Tsc2 KO MEFs (k). n = 4, means ± SEM, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. Cd276 promoter activity is suppressed by YY2 knockdown in Tsc2−/− 105K cells (l) and Tsc2 KO MEFs (m). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
YY2 controls CD276 transcription in Tsc2-deficient cells. Enhanced Cd276 promoter activity in Tsc2−/− 105K cells expressing empty vector (EV) compared to reconstitution of TSC2 (a) and in Tsc2 KO MEFs compared to Tsc2-WT MEFs (b). Relative luciferase activity was determined by a dual-luciferase assay system. psiCHECK2-Cd276 encodes the Cd276 promoter. Empty psiCHECK2 vector was used as the negative control. n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Raptor, mTOR or S6K knockdown suppresses Cd276 promoter activity in Tsc2−/− 105K cells (c) and Tsc2 KO MEFs (d). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. siRNA knockdown of YY2 reduces B7-H3 protein expression in Tsc2−/− 105K cells (e) and Tsc2 KO MEFs (f) (n = 3). Knockdown of YY2 reduces Cd276 mRNA expression in Tsc2−/− 105K cells (g) and Tsc2 KO MEFs (h). n = 3, means ± SD, two-tailed unpaired Student’s t-test, *p < 0.05, ***p < 0.001. i Promoter region of Cd276 displaying the location of the YY2 binding site. YY2 ChIP-qPCR analysis showing increased YY2 occupancy on the Cd276 promoter in Tsc2−/− 105K cells (j) and Tsc2 KO MEFs (k). n = 4, means ± SEM, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. Cd276 promoter activity is suppressed by YY2 knockdown in Tsc2−/− 105K cells (l) and Tsc2 KO MEFs (m). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
YY2 controls CD276 transcription in Tsc2-deficient cells. Enhanced Cd276 promoter activity in Tsc2−/− 105K cells expressing empty vector (EV) compared to reconstitution of TSC2 (a) and in Tsc2 KO MEFs compared to Tsc2-WT MEFs (b). Relative luciferase activity was determined by a dual-luciferase assay system. psiCHECK2-Cd276 encodes the Cd276 promoter. Empty psiCHECK2 vector was used as the negative control. n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Raptor, mTOR or S6K knockdown suppresses Cd276 promoter activity in Tsc2−/− 105K cells (c) and Tsc2 KO MEFs (d). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. siRNA knockdown of YY2 reduces B7-H3 protein expression in Tsc2−/− 105K cells (e) and Tsc2 KO MEFs (f) (n = 3). Knockdown of YY2 reduces Cd276 mRNA expression in Tsc2−/− 105K cells (g) and Tsc2 KO MEFs (h). n = 3, means ± SD, two-tailed unpaired Student’s t-test, *p < 0.05, ***p < 0.001. i Promoter region of Cd276 displaying the location of the YY2 binding site. YY2 ChIP-qPCR analysis showing increased YY2 occupancy on the Cd276 promoter in Tsc2−/− 105K cells (j) and Tsc2 KO MEFs (k). n = 4, means ± SEM, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. Cd276 promoter activity is suppressed by YY2 knockdown in Tsc2−/− 105K cells (l) and Tsc2 KO MEFs (m). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
B7-H3 expression is regulated by mTORC1.B7-H3 protein expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (a), TSC2−/− 621-101 angiomyolipoma-derived cells with reconstitution of TSC2 or empty vector (EV) (b), and Tsc2-WT and Tsc2 KO MEFs (c). For all figure legends, n = 3 indicates representative of 3 biologic samples (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells with stable reconstitution of TSC2 or empty vector (EV) (d), TSC2−/− 621-101 angiomyolipoma tumor cells with reconstitution of TSC2 or empty vector (EV) (e), Tsc2-WT and Tsc2 KO MEFs (f). Means ± SD, two-tailed unpaired Student’s t-test (d, f) or one-way ANOVA with Dunnett’s multiple comparisons test (e), *p < 0.05, **p < 0.01. n = 3. B7-H3 protein expression in Tsc2−/− 105K cells (g), TSC2−/− 621-101 angiomyolipoma tumor cells (h), and Tsc2 KO MEFs (i) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1 or vehicle for 24 hr (n = 3). B7-H3 mRNA expression in Tsc2−/− 105K cells (j), TSC2−/− 621-101 angiomyolipoma tumor cells (k), and Tsc2 KO MEFs (l) treated with 20 nM rapamycin (Rapa), 500 nM Torin 1, or vehicle for 24 hr. n = 3, means ± SD, one-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001. m Immunoblotting analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 h hr (n = 3). n qRT-PCR analysis of Tsc2-WT and Tsc2 KO MEFs transfected with non-targeting control siRNA (Ctrl) or SMARTpool siRNAs targeting either Raptor, Rictor, mTOR, S6K, or 4E-BP1 for 48 hr. n = 3, means ± SD, two-way ANOVA with Dunnett’s multiple comparisons test, ***p < 0.001, ****p < 0.0001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse B7-H3 by Western Blot
YY2 controls CD276 transcription in Tsc2-deficient cells. Enhanced Cd276 promoter activity in Tsc2−/− 105K cells expressing empty vector (EV) compared to reconstitution of TSC2 (a) and in Tsc2 KO MEFs compared to Tsc2-WT MEFs (b). Relative luciferase activity was determined by a dual-luciferase assay system. psiCHECK2-Cd276 encodes the Cd276 promoter. Empty psiCHECK2 vector was used as the negative control. n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Raptor, mTOR or S6K knockdown suppresses Cd276 promoter activity in Tsc2−/− 105K cells (c) and Tsc2 KO MEFs (d). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. siRNA knockdown of YY2 reduces B7-H3 protein expression in Tsc2−/− 105K cells (e) and Tsc2 KO MEFs (f) (n = 3). Knockdown of YY2 reduces Cd276 mRNA expression in Tsc2−/− 105K cells (g) and Tsc2 KO MEFs (h). n = 3, means ± SD, two-tailed unpaired Student’s t-test, *p < 0.05, ***p < 0.001. i Promoter region of Cd276 displaying the location of the YY2 binding site. YY2 ChIP-qPCR analysis showing increased YY2 occupancy on the Cd276 promoter in Tsc2−/− 105K cells (j) and Tsc2 KO MEFs (k). n = 4, means ± SEM, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. Cd276 promoter activity is suppressed by YY2 knockdown in Tsc2−/− 105K cells (l) and Tsc2 KO MEFs (m). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Source data and exact p values are provided in the Source data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36869048), licensed under a CC-BY license. Not internally tested by R&D Systems.
