Skip to main content
Fig. 3 | Cancer Cell International

Fig. 3

From: Molecular mechanism leading to SAHA-induced autophagy in tumor cells: evidence for a p53-dependent pathway

Fig. 3

TP53 gene transfer restores apoptosis induction by SAHA in ESS-1 cells. a The expression vector pCMV6-XL5 containing the CMV promoter, either wild-type (TP53-WT; upper panel) or the constructed 637C>T mutated (TP53-637C>T; lower panel) human full-length cDNA for TP53, and the signal sequence for the human growth hormone signal (hGH polyA). b Detection of p53 expression by Western blot in ESS-1 cells transiently transfected with the TP53-WT or -637C>T expression vectors. MES-SA cells were used as a positive control. β-tubulin was used as loading control. c Transient transfection of ESS-1 cells with TP53-WT and TP53-637C>T expression constructs shown in a. Following 3 µM SAHA treatment, TP53-WT-transfected cells underwent close to complete lysis 24 h after transfection (upper middle panel) while no obvious effects could be seen in SAHA-treated (upper panel) or –untreated (lower panel) ESS-1 (left panel) and TP53-637C>T transfected (right panel) control cells. (Magnification: 20-fold) d Screening for p53-induced mitochondria-mediated apoptosis by Western blot analysis. Cell extracts of ESS-1 cells that were untreated (U) or transiently transfected with TP53-WT, or TP53-637C>T expression vectors and were treated or non-treated (CO; 6 h) with 3 µM SAHA were electrophoresed by SDS PAGE, blotted, and monitored after 6 or 12 h with antibodies against the cleaved pro-apoptotic protein PUMA (23 kDa), caspase-9 (CASP-9; 35/37 kDa), and cleaved PARP-1 (89 kDa). β-tubulin was used as loading control

Back to article page