Figure 6

Combined ChIP-MeDIP analysis reveals changes in methylation and Pol II levels at a pro-inflammatory gene locus in diabetic mouse kidney. Sheared cross-linked kidney chromatin from two-month-old diabetic OVE26 (on FVB background) (A), and two-month-old leptin resistant insulin resistant obese C57BL/6J ob/ob mice (B) and their respective normal age-matched controls were assayed using anti-5mC, Pol II CTD and H3K9,14Ac antibodies. The same cross-linked chromatin samples were used for both MeDIP and ChIP assays. MeDIP and ChIP DNA were analyzed at the indicated sites of the MCP-1 and TGF-β1 genes in real-time PCR. Data represent mean ± SEM (6 animals from each strain), expressed either as % Input (panels 1-3) or 5mC/Pol II ratio (panel 4).