Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF) induced the claudin-5 expression in hCMEC/D3 cells by activating the PI3K/AKT/FOXO1 pathway. Effects of US-CM and GDNF on the phosphorylated FOXO1 (p-FOXO1)/FOXO1 ratio, total FOXO1 expression (A). Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF)-induced VE-cadherin expression in hCMEC/D3 cells by activating the PI3K/AKT/ETS1 and MAPK/ERK/ETS1 pathways. Effects of US-CM and GDNF on total (A) and nuclear (B) ETS1 expression. Effects of LY and U0 on 200 pg/ml GDNF-induced total (C) and nuclear (D) ETS1 expression. Expression levels of total (E) and the nuclear ETS1 (F) in hCMEC/D3 cells after knocking down ETS1 with siRNA (siETS1). (G) Effects of GDNF and siETS1 on the expression of VE-cadherin and claudin-5. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test. Figure 5—source data 1.The western blot raw images in Figure 5.Figure 5—source data 2.The labeled western blot images in Figure 5.Figure 5—source data 3.Excel file containing summary data and data analysis of Figure 5.The western blot raw images in Figure 5.The labeled western blot images in Figure 5.Excel file containing summary data and data analysis of Figure 5. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF)-induced claudin-5 & VE-cadherin expression in hCMEC/D3 by activating the PI3K/AKT & MAPK/ERK signaling.(A) Effects of 3 μM LY294002 (LY) on levels of claudin-5, VE-cadherin, & p-AKT/AKT in hCMEC/D3 stimulated by 200 pg/ml GDNF. (B) Effects of 2 μM U0126 (U0) on levels of claudin-5, VE-cadherin, & p-ERK/ERK in hCMEC/D3 stimulated by 200 pg/ml GDNF. (C) Effects of 5 μM SP600125 (SP) on levels of claudin-5, VE-cadherin, & p-JNK/JNK in hCMEC/D3 stimulated by 200 pg/ml GDNF. (D) Effects of 2 μM SB203580 (SB) on levels of claudin-5, VE-cadherin, & p-p38/p38 in hCMEC/D3 stimulated by 200 pg/ml GDNF. (E) Effects of anti-GDNF antibody on GDNF-induced p-AKT/AKT & p-ERK/ERK ratios. (F) Effects of 3 μM LY on levels of claudin-5, VE-cadherin, & p-AKT/AKT in hCMEC/D3 stimulated by US-CM. (G) Effects of 2 μM U0 on levels of claudin-5, VE-cadherin, & p-ERK/ERK in hCMEC/D3 stimulated by US-CM. (H) Effects of 5 μM SP on levels of claudin-5, VE-cadherin, & p-JNK/JNK in hCMEC/D3 stimulated by US-CM. (I) Effects of 2 μM SB on levels of claudin-5, VE-cadherin, & p-p38/p38 in hCMEC/D3 stimulated by US-CM. (J) Effects of anti-GDNF antibody on US-CM-induced p-AKT/AKT & p-ERK/ERK ratios. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate per biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test or Welch’s ANOVA test. Figure 3—source data 1.The WB raw images in Figure 3.Figure 3—source data 2.The labeled WB images in Figure 3.Figure 3—source data 3.Excel file containing summary data & data analysis of Figure 3.The WB raw images in Figure 3.The labeled WB images in Figure 3.Excel file containing summary data & data analysis of Figure 3. Image collected & cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
The deficiency of brain glial cell line-derived neurotrophic factor (GDNF) in mice increased the permeability of blood–brain barrier (BBB) and reduced claudin-5 and VE-cadherin expression in mice brains.(A) Experimental configuration of AAV-GFP (shNC) or AAV-shGdnf (shGdnf) intracerebroventricular injection. (B) Effects of brain-specific Gdnf silencing on the expression levels of GDNF, claudin-5, and VE-cadherin in the brains. Effects of brain-specific Gdnf silencing on NaF levels in plasma (C), brain (D), and the ratio of brain to plasma (E). Effects of brain-specific Gdnf silencing on FITC-Dex levels in plasma (F), brain (G), and the ratio of brain to plasma (H). The expression ratios of p-AKT/AKT (I), p-ERK/ERK (J), and p-FOXO1/FOXO1 (K) in the brains of Gdnf silencing mice. (L) The expression level of ETS1 in the brains of Gdnf silencing mice. The above data are shown as the mean ± SEM. Six biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by unpaired t-test, unpaired t-test with Welch’s correction, or Mann–Whitney test. Figure 6—source data 1.The western blot raw images in Figure 6.Figure 6—source data 2.The labeled western blot images in Figure 6.Figure 6—source data 3.Excel file containing summary data and data analysis of Figure 6.The western blot raw images in Figure 6.The labeled western blot images in Figure 6.Excel file containing summary data and data analysis of Figure 6. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF) induced the claudin-5 expression in hCMEC/D3 cells by activating the PI3K/AKT/FOXO1 pathway. (G) Effects of LY on the claudin-5 expression upregulated by siFOXO1. The above data are shown as the mean± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test, Welch’s ANOVA test, or Kruskal–Wallis test. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF) induced the claudin-5 expression in hCMEC/D3 cells by activating the PI3K/AKT/FOXO1 pathway. The expression levels of claudin-5, and VE-cadherin (D) in hCMEC/D3 cells transfected with FOXO1 siRNA (siFOXO1). NC: negative control. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Neurons and astrocytes upregulated claudin-5 and VE-cadherin expression in hCMEC/D3 cells due to glial cell line-derived neurotrophic factor (GDNF) secretion. Effects of 3 μM RET tyrosine kinase inhibitor SSP-86 (SPP), and 5 μM Src family kinases inhibitor PP2 on the upregulation of claudin-5 and VE-cadherin induced by 200 pg/mL GDNF (K). Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Neurons and astrocytes upregulated claudin-5 and VE-cadherin expression in hCMEC/D3 cells due to glial cell line-derived neurotrophic factor (GDNF) secretion. Effects of TGF-beta (G) on the expression of claudin-5 and VE-cadherin. The dosages have been marked in the figure. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF)-induced claudin-5 & VE-cadherin expression in hCMEC/D3 by activating the PI3K/AKT & MAPK/ERK signaling.(A) Effects of 3 μM LY294002 (LY) on levels of claudin-5, VE-cadherin, & p-AKT/AKT in hCMEC/D3 stimulated by 200 pg/ml GDNF. (B) Effects of 2 μM U0126 (U0) on levels of claudin-5, VE-cadherin, & p-ERK/ERK in hCMEC/D3 stimulated by 200 pg/ml GDNF. (C) Effects of 5 μM SP600125 (SP) on levels of claudin-5, VE-cadherin, & p-JNK/JNK in hCMEC/D3 stimulated by 200 pg/ml GDNF. (D) Effects of 2 μM SB203580 (SB) on levels of claudin-5, VE-cadherin, & p-p38/p38 in hCMEC/D3 stimulated by 200 pg/ml GDNF. (E) Effects of anti-GDNF antibody on GDNF-induced p-AKT/AKT & p-ERK/ERK ratios. (F) Effects of 3 μM LY on levels of claudin-5, VE-cadherin, & p-AKT/AKT in hCMEC/D3 stimulated by US-CM. (G) Effects of 2 μM U0 on levels of claudin-5, VE-cadherin, & p-ERK/ERK in hCMEC/D3 stimulated by US-CM. (H) Effects of 5 μM SP on levels of claudin-5, VE-cadherin, & p-JNK/JNK in hCMEC/D3 stimulated by US-CM. (I) Effects of 2 μM SB on levels of claudin-5, VE-cadherin, & p-p38/p38 in hCMEC/D3 stimulated by US-CM. (J) Effects of anti-GDNF antibody on US-CM-induced p-AKT/AKT & p-ERK/ERK ratios. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate per biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test or Welch’s ANOVA test. Figure 3—source data 1.The WB raw images in Figure 3.Figure 3—source data 2.The labeled WB images in Figure 3.Figure 3—source data 3.Excel file containing summary data & data analysis of Figure 3.The WB raw images in Figure 3.The labeled WB images in Figure 3.Excel file containing summary data & data analysis of Figure 3. Image collected & cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
The contribution of VE-cadherin on the glial cell line-derived neurotrophic factor (GDNF)-induced claudin-5 expression. Effects of the VE-cadherin siRNA (siVE-Cad) on mRNA expression of VE-cadherin (A) and claudin-5 (B). Effects of siVE-Cad and GDNF on claudin-5 and VE-cadherin protein expression (C). NC: negative control plasmids. The above data are shown as the mean ± SEM. Four biological replicates per group. Two technical replicates for A and B and one technical replicate for C. Statistical significance was determined using unpaired t-test or one-way ANOVA test followed by Fisher’s LSD test. Figure 4—figure supplement 1—source data 1.The western blot raw images in Figure 4—figure supplement 1.Figure 4—figure supplement 1—source data 2.The labeled western blot images in Figure 4—figure supplement 1.Figure 4—figure supplement 1—source data 3.Excel file containing summary data and data analysis of Figure 4—figure supplement 1.The western blot raw images in Figure 4—figure supplement 1.The labeled western blot images in Figure 4—figure supplement 1.Excel file containing summary data and data analysis of Figure 4—figure supplement 1. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF)-induced claudin-5 & VE-cadherin expression in hCMEC/D3 by activating the PI3K/AKT & MAPK/ERK signaling.(A) Effects of 3 μM LY294002 (LY) on levels of claudin-5, VE-cadherin, & p-AKT/AKT in hCMEC/D3 stimulated by 200 pg/ml GDNF. (B) Effects of 2 μM U0126 (U0) on levels of claudin-5, VE-cadherin, & p-ERK/ERK in hCMEC/D3 stimulated by 200 pg/ml GDNF. (C) Effects of 5 μM SP600125 (SP) on levels of claudin-5, VE-cadherin, & p-JNK/JNK in hCMEC/D3 stimulated by 200 pg/ml GDNF. (D) Effects of 2 μM SB203580 (SB) on levels of claudin-5, VE-cadherin, & p-p38/p38 in hCMEC/D3 stimulated by 200 pg/ml GDNF. (E) Effects of anti-GDNF antibody on GDNF-induced p-AKT/AKT & p-ERK/ERK ratios. (F) Effects of 3 μM LY on levels of claudin-5, VE-cadherin, & p-AKT/AKT in hCMEC/D3 stimulated by US-CM. (G) Effects of 2 μM U0 on levels of claudin-5, VE-cadherin, & p-ERK/ERK in hCMEC/D3 stimulated by US-CM. (H) Effects of 5 μM SP on levels of claudin-5, VE-cadherin, & p-JNK/JNK in hCMEC/D3 stimulated by US-CM. (I) Effects of 2 μM SB on levels of claudin-5, VE-cadherin, & p-p38/p38 in hCMEC/D3 stimulated by US-CM. (J) Effects of anti-GDNF antibody on US-CM-induced p-AKT/AKT & p-ERK/ERK ratios. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate per biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test or Welch’s ANOVA test. Figure 3—source data 1.The WB raw images in Figure 3.Figure 3—source data 2.The labeled WB images in Figure 3.Figure 3—source data 3.Excel file containing summary data & data analysis of Figure 3.The WB raw images in Figure 3.The labeled WB images in Figure 3.Excel file containing summary data & data analysis of Figure 3. Image collected & cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF)-induced VE-cadherin expression in hCMEC/D3 cells by activating the PI3K/AKT/ETS1 and MAPK/ERK/ETS1 pathways. Effects of US-CM and GDNF on total (A) and nuclear (B) ETS1 expression. Effects of LY and U0 on 200 pg/ml GDNF-induced total (C) and nuclear (D) ETS1 expression. Expression levels of total (E) and the nuclear ETS1 (F) in hCMEC/D3 cells after knocking down ETS1 with siRNA (siETS1). (G) Effects of GDNF and siETS1 on the expression of VE-cadherin and claudin-5. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test. Figure 5—source data 1.The western blot raw images in Figure 5.Figure 5—source data 2.The labeled western blot images in Figure 5.Figure 5—source data 3.Excel file containing summary data and data analysis of Figure 5.The western blot raw images in Figure 5.The labeled western blot images in Figure 5.Excel file containing summary data and data analysis of Figure 5. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF)-induced VE-cadherin expression in hCMEC/D3 cells by activating the PI3K/AKT/ETS1 and MAPK/ERK/ETS1 pathways. Effects of US-CM and GDNF on total (A) and nuclear (B) ETS1 expression. Effects of LY and U0 on 200 pg/ml GDNF-induced total (C) and nuclear (D) ETS1 expression. Expression levels of total (E) and the nuclear ETS1 (F) in hCMEC/D3 cells after knocking down ETS1 with siRNA (siETS1). (G) Effects of GDNF and siETS1 on the expression of VE-cadherin and claudin-5. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test. Figure 5—source data 1.The western blot raw images in Figure 5.Figure 5—source data 2.The labeled western blot images in Figure 5.Figure 5—source data 3.Excel file containing summary data and data analysis of Figure 5.The western blot raw images in Figure 5.The labeled western blot images in Figure 5.Excel file containing summary data and data analysis of Figure 5. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Neurons and astrocytes upregulated claudin-5 and VE-cadherin expression in hCMEC/D3 cells due to glial cell line-derived neurotrophic factor (GDNF) secretion. Effects of GDNF (D) on the expression of claudin-5 and VE-cadherin. The dosages have been marked in the figure. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF) induced the claudin-5 expression in hCMEC/D3 cells by activating the PI3K/AKT/FOXO1 pathway. (E) Effects of FOXO1 overexpression (FOXO1-OE) and GDNF on the expression levels of claudin-5, total FOXO1, and nuclear FOXO1. FOXO1-NC: negative control plasmids. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF)-induced claudin-5 & VE-cadherin expression in hCMEC/D3 by activating the PI3K/AKT & MAPK/ERK signaling.(A) Effects of 3 μM LY294002 (LY) on levels of claudin-5, VE-cadherin, & p-AKT/AKT in hCMEC/D3 stimulated by 200 pg/ml GDNF. (B) Effects of 2 μM U0126 (U0) on levels of claudin-5, VE-cadherin, & p-ERK/ERK in hCMEC/D3 stimulated by 200 pg/ml GDNF. (C) Effects of 5 μM SP600125 (SP) on levels of claudin-5, VE-cadherin, & p-JNK/JNK in hCMEC/D3 stimulated by 200 pg/ml GDNF. (D) Effects of 2 μM SB203580 (SB) on levels of claudin-5, VE-cadherin, & p-p38/p38 in hCMEC/D3 stimulated by 200 pg/ml GDNF. (E) Effects of anti-GDNF antibody on GDNF-induced p-AKT/AKT & p-ERK/ERK ratios. (F) Effects of 3 μM LY on levels of claudin-5, VE-cadherin, & p-AKT/AKT in hCMEC/D3 stimulated by US-CM. (G) Effects of 2 μM U0 on levels of claudin-5, VE-cadherin, & p-ERK/ERK in hCMEC/D3 stimulated by US-CM. (H) Effects of 5 μM SP on levels of claudin-5, VE-cadherin, & p-JNK/JNK in hCMEC/D3 stimulated by US-CM. (I) Effects of 2 μM SB on levels of claudin-5, VE-cadherin, & p-p38/p38 in hCMEC/D3 stimulated by US-CM. (J) Effects of anti-GDNF antibody on US-CM-induced p-AKT/AKT & p-ERK/ERK ratios. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate per biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test or Welch’s ANOVA test. Figure 3—source data 1.The WB raw images in Figure 3.Figure 3—source data 2.The labeled WB images in Figure 3.Figure 3—source data 3.Excel file containing summary data & data analysis of Figure 3.The WB raw images in Figure 3.The labeled WB images in Figure 3.Excel file containing summary data & data analysis of Figure 3. Image collected & cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF) induced the claudin-5 expression in hCMEC/D3 cells by activating the PI3K/AKT/FOXO1 pathway. The expression levels of total and nuclear FOXO1 (C) in hCMEC/D3 cells transfected with FOXO1 siRNA (siFOXO1). NC: negative control. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF)-induced VE-cadherin expression in hCMEC/D3 cells by activating the PI3K/AKT/ETS1 and MAPK/ERK/ETS1 pathways. Effects of US-CM and GDNF on total (A) and nuclear (B) ETS1 expression. Effects of LY and U0 on 200 pg/ml GDNF-induced total (C) and nuclear (D) ETS1 expression. Expression levels of total (E) and the nuclear ETS1 (F) in hCMEC/D3 cells after knocking down ETS1 with siRNA (siETS1). (G) Effects of GDNF and siETS1 on the expression of VE-cadherin and claudin-5. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test. Figure 5—source data 1.The western blot raw images in Figure 5.Figure 5—source data 2.The labeled western blot images in Figure 5.Figure 5—source data 3.Excel file containing summary data and data analysis of Figure 5.The western blot raw images in Figure 5.The labeled western blot images in Figure 5.Excel file containing summary data and data analysis of Figure 5. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF) induced the claudin-5 expression in hCMEC/D3 cells by activating the PI3K/AKT/FOXO1 pathway. Effects of US-CM and GDNF on the cytoplasmic p-FOXO1, cytoplasmic FOXO1, and nuclear FOXO1 expression (B). Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GDNF by Western Blot
Glial cell line-derived neurotrophic factor (GDNF)-induced VE-cadherin expression in hCMEC/D3 cells by activating the PI3K/AKT/ETS1 and MAPK/ERK/ETS1 pathways. Effects of US-CM and GDNF on total (A) and nuclear (B) ETS1 expression. Effects of LY and U0 on 200 pg/ml GDNF-induced total (C) and nuclear (D) ETS1 expression. Expression levels of total (E) and the nuclear ETS1 (F) in hCMEC/D3 cells after knocking down ETS1 with siRNA (siETS1). (G) Effects of GDNF and siETS1 on the expression of VE-cadherin and claudin-5. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test. Figure 5—source data 1.The western blot raw images in Figure 5.Figure 5—source data 2.The labeled western blot images in Figure 5.Figure 5—source data 3.Excel file containing summary data and data analysis of Figure 5.The western blot raw images in Figure 5.The labeled western blot images in Figure 5.Excel file containing summary data and data analysis of Figure 5. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/96161), licensed under a CC-BY license. Not internally tested by R&D Systems.
