G9a ehmt2 c6h3

The ChIP assays were performed using a ChIP isolation kit (Millipore, Billerica, MA). We treated 1 × 10⁵ to 5 × 10⁵ murine naïve T cells with 1% formaldehyde to cross-link histones to DNA. The fixed cells were sonicated to yield chromatin fragments of 200–500 base pairs (bp). The antibodies used in the ChIP assays included: H3Ac (Millipore), H4Ac (Upstate Biotechnology, Lake Placid, NY), H3K9Ac (Active Motif, Carlsbad, CA), H3K4Me3 (Millipore), H3K4Me2 (Abcam), H3K4Me (Millipore), H3K9Me2 (Millipore), H3K9Me3 (Abcam) H3K27me2 (Abcam) H3K27me3 (Millipore), p300 (Abcam), PCAF (Abcam), SUV39H1 (Millipore), CBP (Abcam), ESET/SetDB1(Millipore), Ezh2 (Cell Signaling Technology, Beverly, MA), G9a/EHMT2 (C6H3) (Abcam), HP1α (Millipore), HP1β (Active Motif), and HP1γ (Millipore). DNA was recovered by a Chelex 10% slurry and sample boiling method,¹⁰ (link) or using the IP-STAR (Diagenode, Denville, NJ) direct chip protocol followed by the IPPURE DNA purification. 1% Pre-enriched chromatin (input) served as the percentage input for sample quantification, confirmed by fold over IgG changes. The UV-exposed 4% agarose gels were imaged, and the digitized images were analyzed using the software VisionWorks (UVP). Protein signals were indexed by measuring their relative mean grey value per area.