Fig. 3: HS-1371 protects cells from TNF-induced necroptosis. | Experimental & Molecular Medicine

Fig. 3: HS-1371 protects cells from TNF-induced necroptosis.

From: HS-1371, a novel kinase inhibitor of RIP3-mediated necroptosis

Fig. 3

a TNF-induced necroptosis was completely blocked by HS-1371 treatment. HT-29 cells were pretreated with four tested inhibitors for 2 h and then treated with TSZ (6 h for immunoblotting, 24 h for cell death assay). Cell lysates were analyzed by immunoblotting, and cell viability was analyzed by MTT assay or phase-contrast microscopy. The results are presented as the means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. b Protection of necroptosis by HS-1371 treatment is similar with dabrafenib, which abolished downstream events. HT-29 cells were pretreated with various necroptosis inhibitors and HS-1371 before TSZ treatment. Cell lysates were analyzed by immunoblotting, and cell viability was analyzed by MTT assay (left panel). Cells were stained with phospho-MLKL antibody and analyzed by confocal fluorescence microscopy (middle panel), and cell death was further analyzed by FACS analysis after PI staining (right panel). c HS-1371 protects cells from TNF-induced necroptosis in a dose-dependent manner. HT-29 cells were pretreated with indicated concentrations of HS-1371 and then treated with TSZ (6 h for immunoblotting, 24 h for cell death assay). Cell lysates were analyzed by immunoblotting, and cell viability was analyzed by MTT assay or phase-contrast microscopy. The results are presented as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001

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