Extra-nuclear signaling of PR to moesin in T47-D cells: ROCK-2.

<p>(A) and (B) T47-D cells were exposed for 15 min to progesterone or MPA (both 100 nM) in the presence or absence of the ROCK-2 inhibitor, Y-27632 (Y - 10 µM) and moesin and phosphorylated moesin were assayed with western analysis. * = P<0.05 vs control, # = P<0.05 vs corresponding progestin. (C) T47-D cells were transfected with scrambled siRNA or ROCK-2 target siRNA for 48 h. ROCK-2 protein expression was detected by western blot. (D) Cells were exposed to 100 nM progesterone or MPA for 15 min after transfection with 100 nM target siRNA for ROCK-2 or scrambled siRNA for 48 h. Cell contents of wild-type or phosphorylated moesin are shown. (E) and (F) Cells were treated with progesterone or MPA (both 100 nM) for 15 min in the presence or absence of ORG 31710 (ORG - 1 µM), of wortmannin (WM - 30 nM), of PTX (100 ng/mL) or of Y-27632 (Y - 10 µM). ROCK-2 was immunoprecipitated with a specific Ab and the IPs were used to phosphorylate the bait protein, myelin basic protein (MBP). ROCK-2 kinase activity is shown as the amount of phosphorylated MBP (P-MBP). * = P<0.05 vs control, # = P<0.05 vs corresponding progestin. The experiments were performed in triplicates and representative images are shown.