Fig. 2
AR differentially regulates the VEGF-A vs. VEGF-C in ccRCC cells. a Protein expressions of AR, VEGF-A, VEGF-C, and VEGF-D were detected by western blot in a series of ccRCC cell lines (SN12-PM6, Caki-1, ACHN, A498, SW839, OSRC-2, 769-P, and 786-O). The prostate cancer cell line C4-2 expressing AR was used as positive control. b Quantitation of protein expressions of AR, VEGF-A, VEGF-C, and VEGF-D in triplicate western blot assays of VHL mutant cell lines (A498, SW839, OSRC-2, 769-P, and 786-O). c Immunoblotting of HIF2α, VEGF-A, VEGF-C, and VEGF-D following 48 h DHT and Enz treatment in SW839 cell line. d Quantitation of protein expressions of AR, VEGF-A, VEGF-C, and VEGF-D in triplicate western blot assay. e The mRNA levels of HIF2α, VEGF-A, and VEGF-C following 24 h DHT and Enz treatment in SW839 cells. f Western blotting of HIF2α, VEGF-A, and VEGF-C in A498 cells with/without AR overexpression (OE-AR). g We used two different sh-RNAs (pLKO1-sh-AR and pLV-sh-AR) to knockdown AR in SW839 cells. The results showed that both sh-RNAs could effectively suppress AR. Moreover, HIF2α and VEGF-A were decreased yet VEGF-C was increased. h, i The mRNA levels of HIF2α, VEGF-A, and VEGF-C in cells with/without AR overexpression in A498 cells (h) and AR knockdown in SW839 cells (i), TBP was used as an internal loading control. Triplicate experiments were carried out to detect mRNA and protein expressions. *p < 0.05, **p < 0.01, were considered statistically significant by t-test
