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. Author manuscript; available in PMC: 2024 Sep 19.
Published in final edited form as: Circulation. 2023 Aug 9;148(12):959–977. doi: 10.1161/CIRCULATIONAHA.123.063332

Figure 6. Suppression of aortic smooth muscle cell (SMC) gene expression by IRF3 via the recruitment of EZH2 and the role of IRF3 in the induction of repressive chromatin remodeling.

Figure 6.

Chromatin immunoprecipitation (ChIP) analysis in cultured human aortic SMCs confirms (A) the dsDNA-induced binding of IRF3 to the promoter region of selected contractile genes (ACTA2, CCN1, MYLK). Treatment with dsDNA increased H3K27me3 modification (B) and EZH2 binding (C) at the promoter of contractile genes in wild-type aortic SMCs but not in STING−/− or IRF3−/− SMCs. D, Silencing EZH2 reversed the dsDNA-induced H3K27me3 modification of selected contractile genes in aortic SMCs. E, Quantitative PCR analysis confirming that EZH2 depletion suppresses the expression of contractile genes. F, Results of a co-immunoprecipitation assay with endogenous proteins from cultured aortic SMCs confirming the EZH2-IRF3 interaction in response to dsDNA treatment. G, Immunofluorescence microscopy showing the colocalization of the individual components of the EZH2-IRF3 complex. H, ChIP assay confirmed that STING and IRF3 are necessary for the binding of EZH2 to the contractile gene promoter. dsDNA, double-stranded DNA. **p=0.0011; ***p<0.001.