Fig. 4.

Role of RIPK1 in mediating IL-1α and HMGB1 secretion in HT-29 cells. a The expression of RIPK1 in the pool targeted cells by western blot. Band densities were measured by densitometric analysis, and expression of RIPK1 were normalised to β-actin. Expression levels were expressed as fold change relative to RIPK1 non targeting control. b Cells were stimulated for 24 h with Poly(I:C) (100 μg/mL). IL-1α and HMGB1 concentrations were determined by ELISA. IL-1α and HMGB1 levels in sgRIPK1-2 and sgRIPK1-3 cells were compared to cells transduced with NTC. Two-way ANOVA with Bonferroni’s correction was used to determine statistical differences. c Cells transduced with sgRNAs were stimulated for 24 h with Poly(I:C). MTT absorbance was measured. Cell viability of stimulated cells were compared to unstimulated cells. Bars represent mean data (± SEM). One-way ANOVA with Dunnett’s correction was used to determine statistical differences. *P < 0.05