Fig. 6: Pivotal role of KEAP1-PGAM5-AIFM1 signaling axis in SNG-induced oxeiptosis. | Cell Death Discovery

Fig. 6: Pivotal role of KEAP1-PGAM5-AIFM1 signaling axis in SNG-induced oxeiptosis.

From: Targeting oxeiptosis-mediated tumor suppression: a novel approach to treat colorectal cancers by sanguinarine

Fig. 6

CRC cells were stably transfected with PGAM5-shRNAs. After transfection, cells were treated with SNG (4 μM) for 16 h. Following the treatment, A Western blot analysis of indicated proteins was performed. The signal intensities of western blot bands were normalized to AIFM1 or actin of each group, and fold changes were plotted in a histogram from three independent experiments. Significant difference, **p < 0.01 and ***p < 0.001. B CRC cells were stably transfected with PGAM5-shRNAs. After transfection, cells were treated with the indicated concentration of SNG for 16 h and then cell viability was measured by MTT assay. Data shown are mean ± SD (n = 3) (*p < 0.05, **p < 0.01 and ***p < 0.001) and C crystal violet staining was performed. Scale bar: 10 µm. Indicated cells were stably transfected with AIFM1-shRNA, after transfection, cells were treated with the indicated concentrations of SNG (HT-29 for 16 h and CaCo-2 for 6 h), respectively. Following the treatment, D and E cell viability was measured by MTT assay. Data shown are mean ± SD (n = 3) (*p < 0.05, **p < 0.01 and ***p < 0.001), and F crystal violet staining was performed. Scale bar: 10 µm. G HT-29 cells were treated with SNG in the presence or absence of NAC for 16 h. Whole cell lysates of treated as well as untreated cells were subjected to immunoprecipitation with the respective antibodies as indicated, followed by detection precipitates (top) and input lysates (bottom) with the appropriate antibodies by Western blotting. HT-29 cells co-treated with H2O2 (250 μM) were exposed to SNG (2 μM) in the presence or absence of NAC. H Western blot analysis of indicated proteins was performed. The signal intensities of western blot bands were normalized to AIFM1 of each group, and fold changes were plotted in a histogram from three independent experiments. Significant difference, *p < 0.05, ***p < 0.001 and ns = no significance, I Cell viability was measured by using MTT assay. Data shown are mean ± SD (n = 3) (*p < 0.05, ***p < 0.001).

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