Histone h2a

Western blot analysis was performed according to previously published methods [23 (link)]. The cell samples were briefly lysed with mammalian protein lysis buffer (Thermo Fisher Scientific) containing a protease inhibitor mixture (Roche Applied Science, Switzerland). Twenty micrograms of total protein was separated by 10% (wt/vol) SDS-PAGE and transferred onto a nylon membrane (Merck Millipore, Germany). After being blocked with 2% (wt/vol) bovine serum albumin (Sigma-Aldrich, Saint Louis, MO 63103, USA), the antibodies were then used to probe the membrane. The following antibodies were used: histone H2A (CST, 12349, Danvers, Massachusetts, USA) 1 : 1000, LDLR (Millipore, MABS26) 1 : 1000, PCNA (CST, 13110) 1 : 1000, PCSK9 (Circulex, CY-P1037, MBL, Japan) 1 : 1000-2000, β-actin (CST, 4970) 1 : 1000, GAPDH (CST, 5174) 1 : 1000, Akt (pan) (CST, 4691) 1 : 1000, phospho-Akt (Ser473) (CST, 4060) 1 : 1000, p44/42 MAPK (Erk1/2) (CST, 4695) 1 : 1000, and phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (CST, 4370) 1 : 1000. The secondary antibodies used in the western blotting are as follows: anti-mouse IgG, HRP (CST, 7076) and anti-rabbit IgG, HRP (CST, 7074). The protein expression in the western blot analysis was measured by gray analysis software (ImageJ).

The following antibodies were used for Western blotting, immunoprecipitations and chromatin immunoprepicitation: mouse monoclonal antibodies against HA.11 (16B12, Covance), Flag (M2 and M5, Sigma), Xpress (R910-25, Invitrogen), alpha-tubulin (B-5-1-2, Sigma), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH; 6C5, Millipore), Beta Actin (Sigma, A5441), BRG1 (PA5-17008, Thermo Scientific), LaminB (sc-6216, Santa Cruz), RBPJ (Hyb-T6709, Cosmo Bio Co), CtBP1 (Q13363, Millipore), WDR48 (HPA038421, SIGMA or rabbit polyconal sera, a kind gift of Alan D’Andrea), WDR20 (A301-657A, Bethyl Laboratories), USP46 (HPA007288, Sigma), p16 (clone JC8, sc-56330, Santa Cruz Biotechnology), NF-kB p65 (8242, Cell Signalling), EBNA1 (13-156-100, Advanced Biotechnologies), EBNA2 (PE2, [80 (link)]), EBNA3A (F115P, Exalpha Biologicals), EBNA3B (F120P, Exalpha Biologicals), EBNA3C (A10, [81 (link)]), LMP1 (S12, [82 (link)]), EBNALP (4D3, a kind gift of Yasushi Kawaguchi [83 (link)]). histone H2A (#07–146, Millipore), Ub-histone H2A (#05–678, Millipore), histone H2B (07–371, Millipore), Ub-histone H2B (#07–371, Millipore), PHLPP1 (A300-660A, Bethyl), PHLPP2 (A300-661A, Bethyl), Akt (#4691, Cell signaling), p-Akt (#4060, Cell signaling), and GFP (sc-5384, Santa Cruz).

Whole cell extracts for western blotting were prepared as described previously (17 (link)). Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and transferred onto Immobilon-FL PVDF membranes (Millipore) according to standard procedures. Membranes were probed with the following antibodies: APE1 (Novus Biologicals, NB100–101), XRCC1 (Thermo Scientific, MS-1393-P0), α-actin (Abcam, ab6276), p21 (Cell Signaling Technology, 2947), p53 (Santa Cruz, sc-126), PAR (Trevigen, 4335-AMC-050), histone H2A.X phosphorylated at Ser139 (γH2AX, 05–636, Millipore), Sp1 (Millipore, 07–645), histone H3 (Cell Signaling Technology, 4499). Secondary antibodies conjugated with Alexa Fluor 680 (Molecular Probes) and IRDye^® 800 (Rockland) fluorescent dyes were used. Detection and quantification was carried out using an Odyssey image analysis system (Li-Cor Biosciences).