Fig. 2: RIP3-independent sensitization of TRAIL-induced cell death by MLKL depletion. | Cell Death & Disease

Fig. 2: RIP3-independent sensitization of TRAIL-induced cell death by MLKL depletion.

From: Reduction in MLKL-mediated endosomal trafficking enhances the TRAIL-DR4/5 signal to increase cancer cell death

Fig. 2

a HT-29 cells expressing MLKL shRNA, or non-silencing control were analyzed by western blotting (left panel). These cells were treated with varying doses of TRAIL for 48 h and cell viability was analyzed by MTT assay (right panel). The results are presented as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. b HT-29 cells stably expressing MLKL shRNA or a non-silencing control were treated with TRAIL (50 ng/ml) in time-dependent manner. The cells were harvested, and total lysates were analyzed by western blotting. c HT-29 cells expressing RIP3 shRNA, MLKL shRNA, or a non-silencing control were analyzed by western blotting (left panel), and these cells were treated with varying doses of TRAIL for 48 h. Cell viability was analyzed by MTT assay (right panel). The results are presented as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. d HT-29 cells stably expressing MLKL shRNA or a non-silencing control were treated with TRAIL (20 ng/ml) + SMAC mimetic (200 nM) + zVAD (20 μM) (left panel). HT-29 cells stably expressing RIP3 shRNA, MLKL shRNA or non-silencing control were treated with TNF (30 ng/ml) + SMAC mimetic (200 nM) + zVAD (20 μM) (right panel) for 24 h. Cell viability was analyzed by MTT assay. The results are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. e RIP3-expressing HeLa cells expressing MLKL shRNA, or a non-silencing control were analyzed by western blotting, and these cells were treated with varying doses of TRAIL for 12 h and cell viability was analyzed by MTT assay (upper panel) or phase-contrast microscopy (bottom panel). The results are presented as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. Scale bars, 100 μm. f RIP3-expressing HeLa cells expressing MLKL shRNA, or a non-silencing control were treated with varying doses of TRAIL for 5 h. The cells lysates were analyzed by western blotting. g Cells from (f) were treated with TNF + or TRAIL + SMAC mimetic + zVAD for 24 h. Cell viability was analyzed by MTT assay (left panel) or phase-contrast microscopy (right penal). The results are presented as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. Scale bars, 100 μm.

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