Anti h3k14ac

An EZ-Magna ChIP™ kit (cat. no. 17-371; Sigma-Aldrich; Merck KGaA) was used for ChIP. Chromatin was immunoprecipitated for 24 h at 4°C using anti-HDAC2 (cat. no. ab124974; 1 µg; Abcam), anti-H3K14ac (cat. no. ab203952; 1 µg; Abcam) and anti-c-Myc (cat. no. 18583S; 1 µg; Cell Signaling Technology, Inc.) antibody. Rabbit IgG immunoprecipitation was used as a negative control. 1/100 of total cell lysate was used as an internal control. qPCR was used to analyze precipitated DNA using specific primers (Table II) and it was performed as described above. The signals were calculated as the percentage of input.

HCT116, HeLa, HEK293T, A549, H1299, Calu3 and MDA-MB-231 cells were cultured in McCoy’s 5A medium or DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a 37°C incubator with a 5% CO₂ and humidified atmosphere. The antibodies used in this study include: anti-H1 and anti-pan-H3-ac from Active Motif; anti-H1.4, anti-β-actin and anti-Biotin from Santa Cruz; anti-H1K85ac and anti-H1K63ac from PTM Biolabs; anti-H3K14ac, anti-H3K9ac, anti-H4K16ac, anti-histone H3, anti-histone H4 from Abcam; anti-PCAF, anti-phospho-H2AX (S139), anti-H2A, anti-HP1α, anti-SMC1, anti-SUV39H1, anti-pan-acetyl-lysine, anti-HDAC1, anti-HDAC2, anti-HDAC3 and anti-HDAC8 from Cell Signaling; anti-HP1β and anti-HP1γ from GeneTex; anti-FLAG from Sigma; anti-GST from APPLYGEN; anti-GFP from MBL. Adriamycin, etoposide, camptothecin (CPT), isopropy-β-D-thiogalactoside (IPTG), trichostatin A and nicotinamide were purchased from Sigma; anacardic acid was purchased from Selleck.