Anti pik3ip1

Total cell lysates were prepared with 1% triton lysis buffer (25 mM Tris HCl (pH 8.0), 150 mM NaCl, 1% triton-X100, 1 mM dithiothreitol (DTT), protease inhibitor mix (Complete Mini, Roche) and phosphatase inhibitor (PhosphoStop, Roche)) and subjected to SDS-PAGE. The following antibodies were used for immunoblotting: anti-β-actin (Sigma-Aldrich), anti-cleaved PARP (Abcam, #ab32064), anti-PIP4K2A (#5527), anti-PIP4K2B (#9694), anti-cleaved caspase-3 (#9664), anti-pHistone H3(Ser10) (#3377), anti-pAkt(S473) (#9271), anti-pAkt(T308) (#13038), anti-total Akt (#9272), anti-p70S6K(T389) (#9234), anti-total p70S6K (#9202), anti-pErk (#4370), anti-γ-Histone H2AX (#9718) (Cell signaling), and anti-PIK3IP1 (Proteintech, #16826-1-AP). The secondary antibodies used were sheep anti-mouse IgG HRP and donkey anti-rabbit IgG HRP (Amersham; 1:2000 dilution). Immunoreactive proteins were visualized using ECL reagent (Amersham). Where indicated, intensities of protein bands were quantified by densitometry (Odyssey V3.0), normalized to their loading controls and then calculated as fold expression change relative to DMSO control. The uncropped scans of the blots were presented in Supplementary Fig. 9.