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. 2012 Jun 22;1(3):e000901. doi: 10.1161/JAHA.112.000901

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© 2012 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley-Blackwell.

This is an Open Access article under the terms of the Creative Commons Attribution Noncommercial License, which permits use, distribution, and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

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Figure 5. — SRF is required for Notch-mediated SMC differentiation. A and B, Primary human SMCs were treated with TSA or vehicle control dimethyl sulphoxide (DMSO) for 48 h before Western blotting for Jag-1, Dll-4, SRF, and SMA protein levels (A) or quantitative RT-PCR analysis for SRF, myocardin (MYOCD), and MRTFA transcript levels (B). Data are presented as fold change as compared to SMCs treated with DMSO control. Human SMCs were transduced with a nontargeting siRNA (NT) or with siRNA targeted against SRF (si-SRF) for 4 days and were collected to examine the efficiency of knockdown by quantitative RT-PCR (C) and immunoblot (D). E, NT- or siSRF-transduced SMCs were infected with green fluorescent protein (GFP) or Notch1ICD (N1ICD) for 3 days and were collected for analysis of SMC markers. F, SMCs were transduced with GFP or N1ICD with the SM actin promoter reporter construct (SMA-125) or the construct with the mutant CArG box (SMA-125-mutCArG). Cells were treated with TSA or control vehicle DMSO (con) before analysis. Data are presented as fold change compared to SMCs with control DMSO treatment. Data are presented as mean±SD, and asterisks indicate P<0.05. SMA-125 activity was significantly increased with N1ICD compared to GFP, and this activity was significantly reduced with TSA.