Western Blot Analysis of Cellular Proteins

Western blot analysis was performed as described previously [48 (link)]. For whole-cell lysates, cells were lysed in RIPA buffer supplemented with protease and phosphatase inhibitors (both from Roche, Mannheim, Germany). One representative experiment out of at least three performed is shown. Primary antibodies were purchased from Cell Signaling, Frankfurt, Germany [anti-MDM2 (86934; used for all western blots); anti-p21 (2947); anti-PML (33156; used for all western blots)], Santa Cruz Biotechnology [anti-p53 (DO-1, used for all western blots except for blots shown in Figs. 5B, 6B); anti-p53 (FL393, for Figs. 5B, 6B), BD Transduction Laboratories, Heidelberg, Germany [anti-Bax (610982)], Merck Millipore, Darmstadt, Germany [anti-DR4 (AB16955)], Epitomics, California, USA [α-Tubulin (1878-1)], ProScience Incorporated, Poway, USA [anti-TRAIL-R2 (2019)] and from Sigma-Aldrich [anti-β-Actin (A5441)]. Secondary antibodies were purchased from Cell Signaling [anti-rabbit-HRP (7074) and anti-mouse-HRP (7076)].