Anti h3k18ac

For qChIP assays in cells, Mefs were incubated with 1% formaldehyde/1% paraformaldehyde for 5 min followed by the addition of 125 mM Glycine to stop the reaction. Cells were then washed in phosphate-buffered saline, resuspended in lysis buffer (10 mM Tris pH 8, 140 mM NaCl, 0.1% SDS, 0.5% Triton X-100, 0.05% NaDoc, 1 mM EDTA, 0.5 mM EGTA, and protease inhibitors) and chromatin was sheared by sonication (epishear, Active motif). qChIPs were carried out by incubating chromatin (Input) with protein G-Dynabeads and the different antibodies: control (irrelevant IgG), affinity-purified rabbit anti-E4F1 polyclonal^{21 (link)}, anti-p53 (1C12, Cell Signaling, 10 μl per IP), anti-H3K4me3 (Cell Signaling, 10 μl per IP), anti-H3H9me3 (Cell Signaling, 10 μl per IP), anti-H3K27me3 (Cell Signaling, 10 μl per IP), anti-H3K18ac (Cell Signaling, 10 μl per IP), anti-H3K27ac (Cell Signaling, 10 μl per IP) antibodies. After overnight incubation, washing, reverse cross-linking, and treatment with both RNase A and Proteinase K, proteins were removed with phenol/chloroform extraction, and DNA was recovered using the NucleoSpin Extract II kit. Input and immunoprecipitated DNA were then analyzed by QPCR using the SYBR Green Master mix on a LightCycler 480 SW 1.5 apparatus (Roche). Results are represented as a percentage of the input. Primers used for qChIP assays are provided in supplementary table 1.

The following antibodies were used as indicated: anti-sirt7 (1:4000, Frdbio) (42 (link)), anti-Myc (1:1000, #sc-40, Santa Cruz Biotechnology), anti-Flag (1:5000, #F1804, Sigma), anti-GAPDH (1:5000, #AC033, ABclonal Technology), anti-HIF-1α (1:1000, #NB100-134, Novus), anti-HIF-1α (1:1000, #36169, Cell Signaling Technology), anti-HIF-2α (1:1000, #NB100-122, Novus), anti-HIF-2α (1:1000, #59973, Cell Signaling Technology), anti-HIF-2α (1:1000, #7096, Cell Signaling Technology), anti-β-actin (1:100,000, #AC026, ABclonal), anti-H3 (1:2000, #A2348, ABclonal), and anti-H3K18Ac (1:1000, #13998, Cell Signaling Technology). In addition, PX478 (#S7612) was purchased from Selleck. MG-132 (#474790) was purchased from Sigma.