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. 2014 Apr 16;28(6):935–948. doi: 10.1210/me.2013-1339

Figure 4.

Figure 4.

Androgen/AR-dependent SGK3 transcription involves ER. A, LNCaP cells were transfected with the siRNA negative control (siNC) or ERβ siRNA (siERβ) for 72 hours. Cell extracts were subjected to Western blotting. B, After being hormone-stripped for 2 days, LNCaP cells were transfected with the siRNA negative control or ERβ siRNA for 48 hours and then were transfected with the luciferase reporters as indicated and cultured in the presence or absence of 1 nM DHT for 24 hours. Luciferase activity of cell extracts was measured and normalized to protein concentration. Data are expressed as means ± SD from 3 independent experiments. **, P < .01, by the Student t test. C, LNCaP cells were transfected with siRNA negative control or ERβ siRNA. Twenty-four hours after transfection, the cells were treated with DMSO or 10 nM DHT for 48 hours. The cells were harvested and subjected to Western blotting with the relevant antibodies. D, LNCaP cells were reversely transfected with siRNA negative control or ERβ siRNA and cultured in the presence of 10 nM DHT for 24 hours and then were transfected with empty vector pCI or ERβ expression vector for 48 hours. Cell extracts were subjected to Western blotting analysis. E, MCF-7 cells were hormone-stripped for 2 days and then were added with or without 10 nM DHT alone or plus 100 nM ICI182,780 (ICI). RT-qPCR was performed to evaluate the SGK3 mRNA level. **, P < .01, by the Student t test. F, MCF-7 cells were transfected with the siRNA negative control or ERα siRNA. Seventy-two hours after transfection, the cells were treated with DMSO or 10 nM DHT for 24 hours. The cells were harvested and subjected to Western blotting with the relevant antibodies. G, After being hormone-stripped for 2 days, MDA-MB-453 cells were transfected with empty vector pSG5 or ERα expression vector for 48 hours and then were transfected with the luciferase reporters as indicated and cultured in the presence or absence of 1 nM DHT for 24 hours. Luciferase activity of cell extracts was measured and normalized to protein concentration. Data are expressed as mean ± SD from 3 independent experiments. n.s., not statistically significant; **, P < .01, by the Student t test. H, MDA-MB-453 cells were hormone-stripped for 2 days and transfected with empty vector or ERα expression vector and then were cultured in the presence or absence of 10 nM DHT for 48 hours. Cell extracts were subjected to Western blotting with the relevant antibodies. NS, nonspecific band(s).