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

Figure 2.

Figure 2

NLRP3 Inflammasome Activation Induces Necrotic Cell Death in the Absence of Caspase-1

(A–F) Control, ASC KO, and CASP1KO THP1 NLRP3 D303N cells were differentiated with PMA for 48 h and then treated with DOX (1 μg/mL). (A) After 6 h, lysates and supernatants were analyzed by western blot. (B) The levels of LDH in the supernatants at the indicated time points were assessed. (C and D) Cells were treated with DOX in the presence of SYTOXG. (C) Merged images of hKO1, SYTOXG, and Hoechst33342 were visualized by confocal microscopy. (D) High-magnification images of DOX-treated CASP1KO THP1 NLRP3 D303N cells. Images were visualized as merged images of fluorescence (right panels) and merged images of fluorescence and bright fields (left panels).

(E–G) Relative fluorescence units of SYTOXG in (E) Control, (F) ASC KO, and (G) CASP1KO THP1 NLRP3 D303N cells were measured at 30-min intervals.

Data are shown as mean ± SD of triplicate (B) or pentaplicate (E–G) of one experiment. (A–G) Data are representative of two independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 as determined by two-way ANOVA with a post hoc test.