H3k4me2 ab7766

Anti-H3K9me2 (ab1220) and H3K4me2 (ab7766) antibodies were purchased from Abcam (Cambridge, UK); anti-E-cadherin (sc-7870) ACTB(sc-47778) and HRP-conjugated secondary (sc-2031, sc-2004) antibodies were purchased from Santa Cruz Biotechnology (Texas, USA). Anti-FLAG (F1804) antibody was obtained from Sigma-Aldrich (St Louis, USA).

MNase-ChIP-seq and MNase-seq protocols were performed according to previous studies^{44 (link)}. In brief, LNCaP cells were exposed to 10 nM DHT or DMSO (Veh) for 4 h. Mono-nucleosomes with solubilized chromatin was achieved by MNase digestion of 2 min at 37 °C, then immunoprecipitated with antibody-conjugated magnetic beads. DNA is phenol extracted and ethanol precipitated. Libraries were prepared from isolated DNA and sent for sequencing on the Illumina HiSeq3000 at the UTHSA sequencing core. All samples were performed in biological replicates. Antibodies include: H3K4me1 (ab8895) 1:500 dilution, H3K4me2 (ab7766) 1:250 dilution, H3K27ac (ab4729) 1:500 dilution, H3K27me3 (ab6002) 1:250 dilution, H3K36me3 (ab9050) 1:250 dilution, H3K79me2 (ab8898) 1:250 dilution from Abcam (Cambridge, MA). H3K4me3 (07-473) 1:500 dilution, H3K9me3 (17-10242) 1:250 dilution from Millipore (Upstate).