Fig. 8.

The PERK–ATF4–CHOP pathway is critically involved in the inhibitory effect of SESN2 on DC pyroptosis. DCs were isolated from the spleens of WT and SESN2^−/− mice treated with or without salubrinal at 24 h after sham surgery or CLP. A, DC pyroptosis was assessed by flow cytometry, and DCs were labeled with active CASP-1 (FAM-FLICA) and 7-AAD. Representative flow cytometry plots are shown on the left, and quantitative analyses of FLICA⁺ 7AAD⁺ DCs are shown on the right (n = 6). B, Immunoblot analysis of the expression of inflammasome markers, including CASP-1, GSDMD, NLRP3, and ASC and signaling molecules, including PERK, ATF4, CHOP, and GRP78. β-actin served as an internal control. C, Representative confocal immunofluorescence microscopy images of NLRP3 colocalized with ASC. DyLight 488 (green)-labeled ASC protein, DyLight 594 (red)-labeled NLRP3 protein, and DAPI (blue)-stained nuclei are shown. The images are representative of six independent samples from each group. The data are presented as the mean ± SD of at least three independent experiments. Statistical significance: ^*P < 0.05 versus the control group. D, Kaplan–Meier survival curve analysis was performed for mice with CLP-induced sepsis for 7 days (n = 12). Mice in the sham group underwent surgery without CLP and received 4 ml/kg PBS at 1 h after the operation, mice in the CLP group received 4 ml/kg PBS at 1 h after CLP, and mice in the CLP + salubrinal group received 20 mg/kg salubrinal at 1 h after CLP