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. Author manuscript; available in PMC: 2009 May 4.
Published in final edited form as: Nat Cell Biol. 2008 May 4;10(6):676–687. doi: 10.1038/ncb1730

Figure 5.

Figure 5

Inhibition of autophagy by cytoplasmic p53. (a, b) PFT-α-mediated stimulation of autophagy in cytoplasts. GFP-LC3-transfected HeLa cells were enucleated by density-gradient centrifugation after cytochalasin B treatment and cultured on polylysine-coated coverslips. Mixtures of cells and cytoplasts (arrows, identified by the lack of Hoechst 33342 staining) were then exposed to three different autophagy inducers: PFT-α, nutrient starvation and rapamycin. Results are means ± s.d. n = 3 experiments). (c-e) Effect of p53 mutants on autophagy in p53-/-HCT116 cells. The p53 domains and mutants used in this study are schematically represented in c. Representative micrographs of cells transfected with plasmids expressing WT, nuclear and cytoplasmic p53 are shown in d. Quantification of GFP-LC3 puncta of p53-/- HCT116 cells transiently transfected with p53 mutants is shown in e (mean ± s.d., n = 3 experiments, *P < 0.05). (f) Detection of the phosphorylation status of AMPKα, AACα, TSC2 and p70S6K in p53-/- HCT116 cells transiently expressing WT or mutant p53 (n = 5).