Fig. 4: SpvB leads to epithelium necroptosis through inhibiting RIPK3 ubiquitination and autophagy-dependent degradation. | Cell Death Discovery

Fig. 4: SpvB leads to epithelium necroptosis through inhibiting RIPK3 ubiquitination and autophagy-dependent degradation.

From: Salmonella pSLT-encoded effector SpvB promotes RIPK3-dependent necroptosis in intestinal epithelial cells

Fig. 4

A, C Caco-2 cells were then infected with WT, ΔspvB, or ΔspvB/pspvB S. Typhimurium strain (MOI of 100) and incubated for 4 h. A RIPK3 levels were determined by RT-qPCR. Values were normalized to those of the housekeeping gene β‐ACTIN and fold‐changes relative to the untreated control were shown. C Cells were harvested and subjected to measurement of caspase-8 activity. B, D–G Caco-2 cells were transiently transfected with pEGFP-N1-SpvB or pEGFP-N1 for 24 h. B Western blot analysis of the expression of RIPK3. D Cells were treated with 5 μM MG132 for 24 h and western blot analysis of the expression of RIPK3. E Cells were treated with 100 nM bafilomycin A1 (Baf A1) and western blot analysis of the expression of RIPK3. F Western blot analysis of the expression of LC3-I, LC3-II, and p62. G Cell lysates were immunoprecipitated with anti-RIPK3 antibody or IgG, then immunoblotted with respective antibodies. 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, **P < 0.005; ns not significant.

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