Human/Mouse TSG-6 Antibody

Detection of TSG-6 by Western Blot

Phosphorylated beta-catenin at S311 by PLK1 induced metastasis in an in vivo model. A549 cells expressing RFP-tagged WT of beta-catenin and WT at S311, S311D, or S311A of beta-cateninmtGSK3 beta were injected intravenously into the tail veins of four-week-old BALB/c nude mice, and the tumorigenic and metastatic properties were evaluated after 15 weeks. A, Representative lung tumors from the mouse model. B, The number of metastatic lung tumors was counted and plotted (n = 5). Data are presented as mean ± SD. C-F, Representative H&E staining (C-D) and Ki-67 staining (E-F) were performed using lung tissue from the mice. The relative density of H&E staining (D) and Ki-67 staining (F) was analyzed and plotted. *p <0.05; **p <0.01; ***p <0.001. Data are presented as mean ± SD. G, Immunoblotting was performed using lung tissue lysates from each mouse model. beta-catenin, RFP, N-cadherin, vimentin, CD44, c-Jun, TSG6, laminin gamma2, and beta-actin were detected using specific antibodies. The band intensity values were quantified using densitometry of Photoshop software, normalized, and plotted (left panel). *p <0.05; **p <0.01; ***p <0.001. H, qRT-PCR was performed for RFP, CDH1, CDH2, CD44, JUN, LAMC2, and TNFAIP6 using lung tissue lysates from each mouse model. The relative mRNA expression was plotted. *p <0.05; **p <0.01; ***p <0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36793862), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of TSG-6 by Western Blot

Clinical relevance of PLK1 and beta-catenin in metastatic NSCLC. A-C, The overall survival (OS) times in NSCLC whole patients (n=1292) (A), NSCLC patients with stage 1 (n=522) (B), and stages 3-4 (n=48) (C) were analyzed according to their PLK1 and CTNNB1 expression levels using KM PLOTTER. High (Hi) and low (Lo) were generated by dividing patients according to their expression at the median cut-off. D, A heatmap analysis was performed from the TCGA dataset of lung adenocarcinoma patients for CTNNB1, PLK1, TNFAIP6, LAMC2, CD44, epithelial marker (CDH1), and mesenchymal markers (CDH2, SNAI1, SNAI2, TWIST1, TWIST2, ZEB1, and ZEB2) in paired normal (left) and tumor tissues (right) depending on stages. E-F, A549 and NCI-H460 (H460) non-small cell lung cancer cells were treated with 2.5 ng/ml of TGF-beta for 48 hours. E, Immunoblotting was performed to measure the protein levels of beta-catenin, laminin gamma2, TSG6, CD44, c-Jun, c-Fos, PLK1, p-PLK1T210, p-Smad2S465/S467, Smad2/3, E-cadherin, N-cadherin, vimentin, SNAI1, SNAI2, and GAPDH in A549 (left panel) and NCI-H460 (H460) cells (right panel). F, The relative band intensities for beta-catenin, laminin gamma2, CD44, TSG6, c-Jun, c-Fos, PLK1, p-PLK1T210, E-cadherin, N-cadherin, vimentin, SNAI1, and SNAI2 were quantified using densitometry of Photoshop software. G, The relative band intensities for beta-catenin/GAPDH, p-PLK1T210/PLK1, and p-Smad2S465/S467/Smad2 were quantified using densitometry of Photoshop software. H-I, qRT-PCR was performed for CTNNB1, CDH1, CDH2, PLK1, TNFAIP6, LAMC2, and CD44 expression in A549 (H) and NCI-H460 (I) cells. Data are presented as mean ± SD of at least three independent experiments (significantly different as compared with experimental control). *p <0.05; **p <0.01; ***p <0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36793862), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of TSG-6 by Western Blot

Phosphorylated beta-catenin at S311 by PLK1 induced metastasis in an in vivo model. A549 cells expressing RFP-tagged WT of beta-catenin and WT at S311, S311D, or S311A of beta-cateninmtGSK3 beta were injected intravenously into the tail veins of four-week-old BALB/c nude mice, and the tumorigenic and metastatic properties were evaluated after 15 weeks. A, Representative lung tumors from the mouse model. B, The number of metastatic lung tumors was counted and plotted (n = 5). Data are presented as mean ± SD. C-F, Representative H&E staining (C-D) and Ki-67 staining (E-F) were performed using lung tissue from the mice. The relative density of H&E staining (D) and Ki-67 staining (F) was analyzed and plotted. *p <0.05; **p <0.01; ***p <0.001. Data are presented as mean ± SD. G, Immunoblotting was performed using lung tissue lysates from each mouse model. beta-catenin, RFP, N-cadherin, vimentin, CD44, c-Jun, TSG6, laminin gamma2, and beta-actin were detected using specific antibodies. The band intensity values were quantified using densitometry of Photoshop software, normalized, and plotted (left panel). *p <0.05; **p <0.01; ***p <0.001. H, qRT-PCR was performed for RFP, CDH1, CDH2, CD44, JUN, LAMC2, and TNFAIP6 using lung tissue lysates from each mouse model. The relative mRNA expression was plotted. *p <0.05; **p <0.01; ***p <0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36793862), licensed under a CC-BY license. Not internally tested by R&D Systems.