Rabbit anti h3k27me3 07449

ChIP analysis was performed as previously reported [27 (link)] by using 5 μg of the following antibodies: rabbit anti-H3K4me3 (07473), rabbit anti-H3K27me3 (07449), rabbit anti-H3AcK9 (07352), or rabbit IgG as a negative control (12307) (all from Millipore, MA, USA). 5 ng of immunoprecipitated DNA and the relative control were used as templates for RT-qPCR analysis, performed in duplicate. Primers utilized to amplify the HNF1/6 consensus on HNF4α promoter are as follows: forward 5′-TCCGAAAGACCCAAGTGTGG-3′ and reverse 5′-GCCAATCACGTCCCAGATCA-3′.

ChIP was performed as described^{39 (link),52 (link)}. Cross-linked chromatin was sheared by sonication and immunoprecipitated using ChIP grade rabbit anti-HA tag (C29F4, Cell Signaling), rabbit anti-histone H3 (ab1791, Abcam), rabbit anti-H3K27me3 (07-449, Millipore), mouse anti-Myc tag (clone 4A6, Millipore) or control mouse or rabbit IgG (Santa Cruz Biotechnology). For qPCR, two independent ChIP samples were analyzed, and each sample was assayed in triplicate using primers that cover the promoter, coding and/or 3′ untranslated regions of KDM6B and NEFM (Table S4). Data were presented as percentage of the input chromatin (bound/input × 100). For ChIP against H3K27me3, data were normalized to histone H3 content obtained by anti-histone H3 ChIP.