Fig. 3: Necroptosis was activated through RIPK1-dependent and RIPK1-independent pathways. | Nature Communications

Fig. 3: Necroptosis was activated through RIPK1-dependent and RIPK1-independent pathways.

From: RIPK1 kinase drove brain microvascular endothelial cells death and blood-brain barrier disruption in neonatal Escherichia coli meningitis

Fig. 3

a, b Immunoblot analysis of cleaved caspase-3, PARP1, GSDMD, and phosphorylated RIPK1, RIPK3, and MLKL in RIPK1 WT and RIPK1 S161N cells (a) or in WT, RIPK1 KO, and RIPK3 KO cells (b) after RS218 infection (MOI = 1). Blots for GAPDH served as loading controls. c Immunoblot analysis of total and phosphorylated levels of RIPK1, RIPK3, and MLKL in WT, TNFR1 KO, TLR4 KO, or double KO hBMECs expressing RIPK1 WT or S161N. Blots for GAPDH served as loading controls. d TLR4 was co-immunoprecipitated with RIPK3 and TRIF in RIPK1 WT and S161N cells. e In RIPK1 S161N background, TLR4 was co-immunoprecipitated with RIPK3 and TRIF in TRIF WT, TRIF KO, and TRIF mRHIM cells. f In the RIPK1 S161N background, immunoblot analysis of total and phosphorylated levels of RIPK1, RIPK3, and MLKL in TRIF WT, TRIF KO, and TRIF mRHIM cells. Blots for GAPDH served as loading controls. All experiments were representative of at least three independent experiments with similar results. Source data are provided as a Source Data file.

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