Figure 5.
Knocking down TrxR influences estrogen responsiveness. MCF-7 cells were transfected with 50 pmol control or TrxR siRNA, treated with ethanol (–E2 and light gray bars) or 10 nM E2 (+E2 and black bars) for 24 h, and processed for protein or mRNA analysis. (A) Proteins were subjected to western analysis with an antibody that recognizes TrxR, PR-A and PR-B, or GAPDH. (B) RNA was isolated and cDNA was synthesized for quantitative RT-PCR analysis with primers specific to TrxR, pS2, PR, cyclin G2, cyclin D1, Bcl2, ERα, and 36B4 (internal control) mRNA sequences. Data from three independent experiments, which had been performed in triplicate, were combined and are presented as the mean±S.E.M. ANOVA was used to detect significant differences in mRNA levels in response to E2 (*P≤0.05) or in response to TrxR siRNA (#P≤0.05).