Anti h4k12ac

Mouse E14 ES cells (a gift from T. Fazzio, University of Massachusetts Medical School, Worcester, MA) were cultured in Dulbecco’s modified Eagle’s medium (Corning) supplemented with 15% (v/v) fetal bovine serum, 1% penicillin/streptomycin (Invitrogen), 1 mM sodium pyruvate (Cellgro), 2 mM l-glutamine (Cellgro), 1% MEM nonessential amino acids (Invitrogen), 55 μM β-mercaptoethanol (Sigma-Aldrich), and mouse leukemia inhibitory factor (10 ng/ml) on gelatin-coated dishes in a 37°C incubator with a humidified, 5% CO₂ atmosphere. The antibodies used in this study include anti-H4K20me2 (Diagenode, C15200205), anti-H4K12ac (Abcam, ab46983), anti-H3K36me3 (Active Motif, 61021), anti-H3K4me3 (Abcam, ab8580), and anti-BrdU (BD Biosciences, 555627). Other antibodies including anti-H3K56ac, anti-H3, anti-H4, anti-H2A, and anti-H2B were generated by our lab using a synthetic peptide or full-length recombinant proteins.

Anti-H4K12ac and anti-H3 antibodies were purchased from Abcam (Paris, France), and anti-BRD2 was from Bethyl Laboratories (Montgomery, TX). Anti-Tip60 antibody was purchased from Upstate Laboratories (Merck Millipore, Darmstadt, Germany). The myc-Suv39H1 expression vector was a kind gift from T. Jenuwein (Max Planck Institute of Immunobiology and Epigenetics, Freiburg, Germany; Peters et al., 2001 (link)).
All siRNAs were purchased from Eurogentec (Angers, France). Sequences were as follows:
Control siRNA: ACUCAAACUCACGAAGGAA-dTdT.
Tip60-1: UGAGAUUGAUGGACGGAAA-dTdT.
Tip60-2: UGAAGAACAUUGAGUGUAU-dTdT.
BRD2 siRNA: GACAAAGGAGGAACUGGCUUUGGAG-dTdT.
The following primers were used:
Tip60 mRNA: 5′-GACCCCTTCCTCTTCTACGT-3′ and 5′-CCGGTCTTCCCTTCTACTTT-3′.
p400 mRNA: 5′-GAAGTTGGCTGCTGCTAAGA-3′ and 5′-CCTCTTCTGGAAACTTTCTG-3′.
Suv39H1 mRNA: 5′-GGGTCCACTTGCATGTTGTAA-3′ and 5′-GGCAACATCTCCCACTTTGT-3′.
Major satellites (expression and ChIP): 5′-GACGACTTGAAAAATGACGAAATC-3′ and 5′-CATATTCCAGGTCCTTCAGTGTGC-3′.
β2m mRNA (expression and ChIP): 5′-CCCGTTCTTCAGCATTTGGA-3′ and 5′- CCGAACATACTGAACTGCTACGTAA-3′.
p53 promoter: 5′-CAGAGCAGAAAGGGACTTGG-3′ and 5′-CTTCACTTGGGCCTTCAAAA-3′.