FIGURE 5.

Foxo1 is a transcriptional activator in the promoter region of GSDMD gene. A series of plasmids containing 5′ unidirectional deletions of the promoter region of the GSMDM gene fused in frame to luciferase gene were transfected into BV2 cell lines as well as pc‐Foxo1. Luciferase assay was performed after 48 hours. A, Dual‐luciferase reporter assay of GSDMD and Foxo1 (N = 3). B, Result of ChIP analysis between GSDMD and Foxo1 (N = 3). C, The mutant GSDMD promoter‐driven luciferase reporters with mutated bases shown in red were constructed, and relative fluorescence intensity was measured (N = 3). D, The mRNA level of GSDMD of BV2 cells after incubated with pc‐Foxo1 in presence with pc‐CD73 or not for 24 hours (N = 3). E, The release of LDH was detected by LDH Assay Kit in different cultured groups. F, The release of IL‐1β, IL‐6, and TNF‐α in different cultured groups measured by ELISA (N = 3). ^** P < .01, ^*** P < .001 versus control; ^# P < .05, ^## P < .01, ^### P < .001 versus pc‐ Foxo1. All data are shown as the mean ± SD independent experiments