Skip to main content
. Author manuscript; available in PMC: 2014 Jun 1.
Published in final edited form as: Steroids. 2013 Feb 1;78(6):542–547. doi: 10.1016/j.steroids.2012.12.014

Figure 3. Effects of MEK inhibition on PR-mediated gene transcription are much greater at 24 hours than at 6 hours.

Figure 3

A) T47D cells were cultured in serum-free medium for 24 hours before pretreatment with DMSO or U0126 (10 μM) for 1 hour, followed by treatment with ethanol or R5020 (10 nM) for 6 hours. PR and tubulin protein expression levels were determined by Western blot analysis. A representative experiment is shown. U0126 does not affect PR expression levels in the absence or presence of R5020. B) In parallel to the experiments in panel A, relative SGK and CCND1 mRNA expression was determined using real-time qPCR and normalized to 18S rRNA expression. SGK gene induction was insensitive to U0126, while CCND1 basal and hormone-mediated gene expression was decreased with U0126. * p<0.05 compared to respective treatment in DMSO group. C) T47D cells were cultured and pretreated as in panel A, followed by treatment with ethanol or R5020 (10 nM) for 24 hours. Relative SGK, NDRG1 and FKBP5 mRNA expression was determined using real-time qPCR and normalized to 18S rRNA expression. SGK gene induction was abrogated by U0126, while NDRG1 gene induction was robustly decreased by U0126. In contrast, FKBP5 gene induction was relatively insensitive to U0126. * p<0.05 compared to DMSO R5020. ****p<0.0001 compared to DMSO R5020.