Phistone h3 s10

Whole cell lysates were prepared in cold SDS lysis buffer (10mM Tris,
2% SDS) supplemented with PhosSTOP phosphatase inhibitor and Complete
protease inhibitor cocktail tablets (Roche) as previously described (27 (link)). The following antibodies were used:
Cdk1, pCdk1 (Y15), GAPDH (Cell Signaling Technology), pHistone H3 (S10), PAR
(Millipore), Wee1 (Santa Cruz Biotechnology), pRPA32 (S4/8) (Bethyl) and RPA32
(Abcam). Immunoblots were quantitated using ImageJ (NIH).

Western blotting was performed and detected as described previously [9 (link), 11 (link)]. Primary antibodies used were polyclonal anti-EGFR (Santa Cruz Biotechnology, TX, USA), monoclonal anti-α-tubulin (Sigma-Aldrich), anti-Lamin B1 (Abcam, MA, USA), Histone H3 (Sigma-Aldrich), p-Histone H3 (S10) (Milipore, MA, USA), Erk1/2, p-Erk1/2 (T202/Y204), AKT, p-AKT (T308), p-EGFR (T669), p-EGFR (Y845), p-EGFR (Y992), p-EGFR (Y998), p-EGFR (Y1045), p-EGFR (S1046/1047), p-EGFR (Y1086), p-EGFR (Y1148), p-EGFR (Y1173) (Cell Signaling Technology, MA, USA). The PDVF membranes were developed using enhanced chemiluminescence (ECL Plus; GE Healthcare Life Sciences, NJ, USA). Subsequently, the films were scanned and analyzed using Image J software (http://rsbweb.nih.gov/ij/).