Protein Profiling by Western Blotting

Western blotting was performed as described previously (Chen et al. 2013b (link); Sun et al. 2014 (link)). Proteins were probed with the following antibodies: SIAH1 rabbit pAb (Abcam, Cambridge, MA, USA), Phospho-p53 (Ser15) rabbit pAb (Cell Signaling, Beverly, MA, USA), p53 mouse mAb (Abcam, Cambridge, MA, USA), β-Actin mouse mAb (Santa Cruz, SantaCruz, CA, USA), Phospho-p38 MAPK (Thr180/Tyr182) rabbit mAb (Cell Signaling, Beverly, MA, USA), p38 MAPK rabbit pAb (Cell Signaling, Beverly, MA, USA), Phospho-MAPKAPK-2 (Thr222) antibody (Cell Signaling, Beverly, MA, USA), MAPKAPK-2 antibody (Cell Signaling, Beverly, MA, USA), PUMA rabbit pAb (Abcam, Cambridge, MA, USA), Bak rabbit pAb (Cell Signaling, Beverly, MA, USA) and Cleaved Caspase-3 (Asp175) rabbit pAb (Cell Signaling, Beverly, MA, USA). The membranes were developed on a Kodak X-OMAT 2000A imaging system (Kodak, Rochester, NY, USA). The intensity of the protein band was analyzed by ImageJ software (1.46b, National Institutes of Health, USA). All Western blot analyses were performed in triplicate.