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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 4. MRTFB was essential for Smad2-mediated VSMC marker expression in NCCs.

Figure 4

(A-B) MRTFB was required for Smad2-mediated VSMC marker gene transcription. Monc-1 cells were transduced with adenovirus expressing GFP (Ad-GFP) or MRTFB shRNA (Ad-shMRTFB) and co-transfected with α-SMA (A) or SM22α (B) promoter construct and pcDNA or Smad2 cDNA as indicated followed by vehicle (Ctrl) or TGF-β treatment (5 ng/ml) for 16 hours. Luciferase assays were performed. Luciferase activity was normalized to renilla activity. *P<0.01 compared to Ad-GFP groups with TGF-β. (C-E) MRTFB was required for Smad2-mediated VSMC marker mRNA and protein expression. Monc-1 cells were treated as in A and B but without promoter transfection. α-SMA (C) and SM22α (D) mRNA expression was detected by qPCR and normalized to GAPDH expression. Ttheir protein expression was detected by western blot (E). *P<0.01 compared to Ad-GFP group treated with TGF-β for both α-SMA (C) and SM22α (D), respectively. (F) Quantitative analysis of protein expression in E by normalized to α-tubulin. *P<0.01 compared to Ad-GFP group for both α-SMA and SM22α, respectively.