Fig. 7: Sestrin2 deficiency exacerbates NLRP3 inflammasome-mediated pyroptosis in cholestatic livers.

A, B HepG2 cells were treated with CDCA (20 to 100 µM) or CA (100 to 500 µM) for 24 h. The release of LDH was measured by an LDH assay kit. C, D HepG2 cells were infected with lentiviruses expressing shRNAs targeting luciferase (sh-Luc) or Sestrin2 (sh-SESN2) and treated with 100 µM CDCA or 500 µM CA for 24 h. The release of LDH was measured by an LDH assay kit. E, F Liver tissues were collected from Sesn2^+/+ Sham, Sesn2^+/+ BDL, Sesn2^−/− Sham, and Sesn2^−/− BDL mice (n = 5–6 mice per group) and analyzed by immunoblotting with the indicated antibodies. GAPDH served as a loading control. Band intensities were quantified and normalized to control band intensities. G qRT–PCR analysis of Nlrp3, Asc, Casp1, and Il-1β mRNA levels in liver tissues from the indicated mice (n = 5–10 mice per group). The data are representative of two (E–G) or three (A–D) independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001; ns not significant (Student’s t test in A, B and two-way ANOVA, followed by Tukey’s (C, D) or Benjamini, Krieger, and Yekutieli’s (E–G) post hoc tests).