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. 2016 Aug 11;6:31509. doi: 10.1038/srep31509

Figure 1. Involvement of p27 and RhoA in the P4-induced enhanced migration in T47D cells.

Figure 1

(A) Treatment with P4 (50 nM) for 4–8 h increased the level of p27 protein in T47D cells. (B) Knock-down of p27 abolished the P4-induced migration enhancement in T47D cells. P4 increased formation of the p27-RhoA complex (C) and membrane translocation of RhoA from the cytosol (D). (E) Pre-treatment with a ROCK inhibitor, Y27632 (5 μM), prevented the P4-induced migration enhancement in T47D cells. For Western blot analyses, data are representative of 2 independent experiments with similar results. Values shown in parentheses represent the quantified results adjusted with G3PDH (A) or with G3PDH and cadherin for the cytosol and the membrane, respectively (D) and expressed as ratio over its own control. cadherin and G3PDH were used as a membranous and cytosolic protein marker, respectively, to confirm the purities of isolation and to verify equivalent sample loading. The gels have been run in the same experimental conditions and the cropped blots were shown. The entire gel pictures of 1C were shown in the Supplemental Fig. 1. In (B,E), values represent the means±s.e.mean. (n = 3). *P < 0.05 different from control group. #P < 0.05 different from P4-treated group. Con, control; IB, immunoblotting; IP, immunoprecipitation; siRNA, small interfering RNA.