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. Author manuscript; available in PMC: 2014 Sep 27.
Published in final edited form as: Circ Res. 2013 Jul 1;113(8):10.1161/CIRCRESAHA.113.301921. doi: 10.1161/CIRCRESAHA.113.301921

Figure 7. MRTFB physically interacted with Smad2 and enhanced Smad2 binding to VSMC marker promoter.

Figure 7

(A-B) Co-IP of Smad2 with MRTFB. Monc-1 cells were transduced with adenovirus expressing Flag-tagged MRTFB for 2 days followed by vehicle (-) or TGF-β treatment (+) for 2 h. Cell lysates were immunoprecipitated with Flag (A) or Smad2 (B) antibody and blotted with Flag or Smad2 antibody as indicated. Input was 5% of the cell lysates. (C) MRTFB enhanced Smad2 binding to SM22α promoter. Monc-1 cells were transduced with adenovirus expressing GFP (Ad-GFP), MRTFB (Ad-MRTFB) or MRTFB shRNA (AdshMRTFB) followed by TGF-β treatment (5 ng/ml) for 2 hours. CHIP was performed as described in Methods. Smad2 binding enrichment was detected by RT-PCR using primers amplifying the functional SBE element in SM22α promoter. (D) Smad2 binding to SM22α promoter was quantified by qPCR, and the binding fold increase was shown. *P<0.001, **P<0.001 compared to Ad-GFP group.