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. 2008 Jun 25;105(26):9047–9052. doi: 10.1073/pnas.0803623105

Fig. 6.

Fig. 6.

HG effect in HVSMC and the role of SUV39H1. (A) IL-6 and MCP-1 mRNA levels in HVSMC grown in NG, mannitol (Man), or HG. RT-qPCR results shown as the percentage of NG. (mean ± SE; *, P < 0.05; **, P < 0.01 vs. NG, n = 3). (B and C) ChIP assay with NG- and HG-treated VSMC using H3K9me3 antibodies. Immunoprecipitated DNA and input DNA was analyzed by PCR with IL-6 promoter specific primers. The bar graph represents relative H3K9me3 levels normalized to input (quantified by densitometry) and shown as the percentage of NG (mean ± SE; #, P < 0.001 vs. NG, n = 3). (D) HVSMC were transfected with EGFP or FLAG-SUV39H1 vectors and inflammatory gene levels measured in total RNA extracted from TNF-α-treated cells (10 ng/ml, 1 h). RT-qPCR results are shown as the percentage of EGFP-transfected cells (mean ± SE of triplicate qPCRs; **, P < 0.01; ***, P < 0.0001 vs. TNF-α-treated EGFP cells; IL-6 n = 3, MCP-1 and MCSF n = 2). (E) HVSMC were transfected with expression plasmids for scrambled control or shSUV39H1 and inflammatory genes quantified from TNF-α-treated cells. RT-qPCR results are shown as TNF-α-induced effect with shSUV39H1, fold over that with control vector (mean ± SE of triplicate qPCRs, *, P < 0.05; #, P < 0.001; ***, P < 0.0001 vs. TNF-α effect in control; IL-6, and MCSF n = 3, MCP-1 n = 2).