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. 2022 Jul 19;26(16):4492–4505. doi: 10.1111/jcmm.17474

FIGURE 3.

FIGURE 3

CRIZO induced pyroptosis and excessive autophagy by accumulation of ROS. (A) HL‐7702 cells were pretreated with or without 10 mM NAC before CRIZO treatment. The levels of ROS were determined by flow cytometric analysis in HL‐7702 cells (n = 3). (B) HL‐7702 cells were treated by 15 μM CRIZO with or without 10 mM NAC for 24 h. The expression levels of cleaved‐caspase1, caspase1‐FL, GSDMD‐N, NLRP3, LC3, mTOR, phosphorylated mTOR and phosphorylated P65 in different groups were determined by Western blotting (n = 3). (C) HL‐7702 cells' caspase1 activity was determined after administration for 24 h (n = 3). (D) The expression levels of P65 in nucleus were determined by Western blotting (n = 3). (E) Nuclear translocation of P65 was observed by immunofluorescence labelling with DAPI (blue) and anti‐P65 (red). Scale bar: 20 μm. (F) Cell viability was measured by CCK8 assay (n = 5). The data are expressed as the mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 vs. vehicle group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. CRIZO 15 μM treatment group.