Anti histone h4ac

The following rabbit antibodies were used for the western blotting experiment, primary antibodies: anti-Histone H4 (Abcam, ab177840, rabbit monoclonal), anti-Histone H4Ac (Millipore, 06-866, rabbit monoclonal), anti-Histone H4K5Ac (Abcam, ab51997, rabbit monoclonal), anti-Histone H4K8Ac (Abcam, ab45166, rabbit monoclonal), anti-Histone H4K16Ac (Abcam, ab109463, rabbit monoclonal), anti-Histone H4K20Ac (Abcam, ab177188, rabbit monoclonal), anti-Histone H4K77Ac (Abcam, ab241117, rabbit polyclonal), anti-Histone H3Ac (Active Motif, 39040, rabbit polyclonal), anti-Histone H2AK5Ac (Active Motif, 39108, rabbit polyclonal), and anti-Histone H2AK9Ac (Active Motif, 39110, rabbit polyclonal). Secondary antibody: goat-anti-rabbit IgG (H + L) (Proteintech, SA00001-2). A dilution of 1:2000 is used for primary and secondary antibodies in the western blotting experiment. For ChIP-seq and ChIP-qPCR experiment: anti-Histone H4Ac (Millipore, 06-866, rabbit monoclonal) is used, with a dilution of 1:200.

ChIP assays were performed as previous described with minor modifications [19 (link)]. The seeds were treated with cross-link buffer (1% formaldehyde) under a vacuum for 1 h. 0.2–0.3 g seeds for each sample were used. The chromatin was extracted and sheared to an average length of 500 bp by sonication, and agarose gel electrophoresis was performed to ensure the proper size of the sheared chromatin. Then the chromatin was immunoprecipitated with anti-histone H3ac (Catalog no 06-599, Millipore, MA, USA), anti-histone H4ac (Catalog no 06-866, Millipore, MA, USA) or anti-HDA15 antibodies [16 (link)], respectively. The cross-linking was then reversed and the amount of each immunoprecipitated DNA fragment was determined by quantitative PCR using gene specific primers (Supplementary Table S1).