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. 2018 May 3;12:57–66. doi: 10.1016/j.omtn.2018.04.016

Figure 6.

Figure 6

miR-125b Promoted α-SMA Expression and Cell Contraction by Enhancing RhoA Activity

(A) miR-125b enhanced RhoA activation. Lysates from JS1 cells that were transfected with the indicated RNA were incubated with the immobilized GST-RBD. The bead-bound proteins were dissolved in 2× Laemmli sample buffer and analyzed by immunoblotting for GTP-bound RhoA. Equal aliquots were used to detect total RhoA in each lysate. (B and C) Knockdown of RhoA, Mrtf-A, or Srf blocked the miR-125b-induced α-SMA expression. JS1 cells were cotransfected with miR-125b and the siRNA targeting RhoA (B), Mrtf-A or Srf (C) for 48 hr, followed by RT-PCR or western blotting analysis. (D) Silencing of RhoA blocked the miR-125b-induced contraction of JS1 cells. Collagen gel containing JS1 cells that were transfected with the indicated RNA was released and then incubated with 10% FBS-containing DMEM for 24 hr before analysis for contraction area. Data are presented as mean ± SEM in (D). *p < 0.05.