Fig. 5

Epigenetic regulation of PAI-1 gene expression by HDAC2. RAW264.7 cells were transfected with siRNA targeting HDAC2 for 48 h, and then stimulated with 100 ng/ml LPS for 2 h. Cells were harvested and subjected to the ChIP assay to analyze the status of acetyl-Histone H3, NFκB p65, and c-Jun at the PAI-1 gene promoter. Immunoprecipitated chromatin was analyzed via qPCR using primers (shown with arrows) targeting the promoter regions of PAI-1. HDAC2 knockdown (a) increased the binding of acetyl-histone H3 to the NFκB p65 and c-Jun binding sites of the PAI-1 gene promoter (b, c) following LPS treatment. HDAC2 knockdown also increased the binding of NFκB p65 and c-Jun to the PAI-1 gene promoter (d, e) after LPS stimulation. Data are expressed as the mean relative expression ± SEM of at least three independent experiments