Figure 7. Extra-nuclear signaling of PR to moesin in T47-D cells: RhoA.

(A) and (B) RhoA activity was assayed in cells treated with progesterone or MPA (both 100 nM) for 15 min in the presence or absence of the pure PR antagonist ORG 31710 (ORG - 1 µM), of the PI3K inhibitor wortmannin (WM - 30 nM), or of the G protein inhibitor, PTX (100 ng/mL). Active, GTP-bound RhoA was immunoprecipitated with Rhoteckin and subsequently assayed with western analysis with an anti-RhoA Ab (lower boxes). The upper boxes show the total RhoA content in the input. * = P<0.05 vs control, # = P<0.05 vs corresponding progestin. (C) and (D) T47-D cells were either mock-transfected or exposed to constitutively active or dominant-negative RhoA (RhoA CA or RhoA DN) and Gα₁₃ (Gα₁₃ CA or Gα₁₃ DN). Cells were then treated with progesterone or MPA (both 100 nM) for 15 min and wild type and P-moesin were analyzed. The experiments were performed in triplicates and representative images are shown.