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. 2022 Feb 2;8:44. doi: 10.1038/s41420-022-00841-9

Fig. 2. SpvB promotes necroptosis of IECs.

Fig. 2

A, C Caco-2 cells were infected with the WT, ΔspvB, or ΔspvB/pspvB S. Typhimurium strain (MOI of 100) and incubated for 4 h. Western blot analysis of the expression of A MLKL and p-MLKL (S358), C RIPK3 and p-RIPK3 (S227). B, D Caco-2 cells were treated with either vehicle (DMSO), B 1 μM necrosulfonamide (NSA) or D 1 µM GSK’872 for 1 h. The cells were then infected with the WT, ΔspvB, or ΔspvB/pspvB S. Typhimurium strain (MOI of 100) and incubated for 24 h. Aliquots of cellular supernatants were subjected to LDH release assay. E HeLa cells were transiently transfected with pCMV-HA-RIPK3 for 24 h. Cells were then infected with WT, ΔspvB, or ΔspvB/pspvB S. Typhimurium strain (MOI of 100) and cultured for 24 h. Aliquots of cellular supernatants were subjected to LDH release assay. F HeLa cells were transiently transfected with pCMV-HA-RIPK3 and pEGFP-N1-SpvB for 24 h. Western blot analysis of the expression of p-MLKL. Data were analyzed with IBM SPSS Statistics 19 and presented as the mean ± SEM using Student’s t-test and ANOVA with S-N-K correction. *P < 0.05; ns not significant.