Autophagy Regulated by Gain of Function Mutant p53 Enhances Proteasomal Inhibitor-Mediated Cell Death through Induction of ROS and ERK in Lung Cancer Cells

Figure 4

Autophagy induction promotes cell death in R273H-P53 cells. (a) Null-P53, wt-P53, and R273H-P53 cells were exposed to varying doses of CQ. After 48 h of exposure, cell viability was measured through MTT assay. (b) CQ-treated R273H-P53 cells were analyzed for apoptosis induction through Annexin V/PI staining followed by flow cytometry. Fold change is represented through bar diagram. (c) R273H-P53 cells were exposed to CQ (10 μM), 3-MA (5 mM), and ALLN (10 μM) for 48 h and immunoblot analysis was performed for LC3B-II and Atg-5. indicates significant difference to untreated control. (d) Cell viability in R273H-P53 cells was measured through MTT assay after 48 h of treatments at various combinations, (d) Rapa (500 nM), Rapa + SS, (e) SS, ALLN+SS, (f) Rapa, ALLN+Rapa, (g) 3-MA, 3-MA+ALLN and represented in the form of bar diagram. (h) Apoptosis was analyzed through flow cytometry using Annexin V/PI staining in R273H-P53 cells, after treatment with various autophagy inducers. Percentage of apoptotic cells is represented through bar diagram. (i) Cell viability was measured through MTT assay in R273H-P53 cells after autophagy inhibition in presence or absence of ALLN. indicates significant difference compared to untreated control, while # indicates significant difference compared to ALLN-treated cells.

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