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. 2020 Apr 18;23(5):101070. doi: 10.1016/j.isci.2020.101070

Figure 5.

Figure 5

Caspase-8 Initiates Gasdermin E Processing during NLRP3 Inflammasome Activation

(A) Pam3CSK4-primed WT and Casp1/11−/− macrophages were pretreated with Z-VAD (20 μM) and then treated with nigericin (5 μM) for 3 h. Lysates and supernatants were analyzed by western blot.

(B–D) Pam3CSK4-primed WT and Casp1/11−/− macrophages were pretreated with DEVD and IETD (20 μM each) and then treated with nigericin. (B) After 6 h, the levels of LDH in the supernatants were assessed. (C) Relative fluorescence units of SYTOXG were measured at 10-min intervals. (D) After 3 h, lysates and supernatants were analyzed by western blot.

(E and F) Control, CASP1 KO, and CASP1 and CASP8 double-KO THP1 cells were differentiated with PMA for 48 h and then treated with nigericin (5 μM) for 8 h (E) LDH release in supernatant was assessed. (F) Lysates and supernatants were analyzed by western blot.

(B, C, and E) Data are shown as mean ± SD of triplicate of one experiment. Data are representative of two (A and D–F) or three (B and C) independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 as determined by two-way ANOVA with a post hoc test. n.s., not significant.