Fig. 8

DCA promotes IL-1β secretion in LPS and ATP-stimulated macrophages. a–d Bone marrow-derived macrophages (BMDMs) isolated from mice were stimulated with LPS plus ATP or DCA alone or together. a Representative images of morphological changes in different treatment groups. Red boxes represent the magnified view (scale bars 100 μm). Red boxes represent the magnified view. The red arrow indicates the pyroptotic cells. b Representative images of PI staining in different treatment groups (scale bars 100 μm). c Quantitative analyses of the PI-positive cells percentage in different groups. d Quantitative analyses of the LDH release percentage in different groups. e–i BMDMs isolated from mice were stimulated with LPS and ATP plus DCA or not. Then, differentially expressed genes (DEGS) were analyzed by RNA sequencing. e The number of DEGS in the LPS and ATP plus DCA group vs. the LPS and ATP group. Red represented upregulated DEGS, and blue downregulated DEGS. f Scatter plot showing DEGS in the LPS and ATP plus DCA group vs. the LPS and ATP group. Genes were plotted based on their expression levels (log10 intensity). Red and green dots represented up- and downregulated genes, respectively. g Protein-protein interaction (PPI) analyses of the upregulated DEGs. Pyroptosis-associated genes were indicated. h KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analyses of the upregulated DEGs. The dot size represents the number of DEGS, and the dot color represents the corresponding p value. i GO (Gene Ontology) pathway enrichment analyses of the upregulated DEGs. The dot size represents the number of DEGS, and the dot color represents the corresponding p value. j Representative immunoblot images of GSDMD-FL, GSDMD-NT, Pro-IL-1β, and Cleaved-IL-1β in different treatment groups. k Relative protein levels of GSDMD-NT/FL and Cleaved-IL-1β/Pro-IL-1β in different treatment groups. The density of protein bands was quantified by Alpha-view software. c, d, and k Data represent means ± SEM; *P < 0.05; **P < 0.01; ***P < 0.001; by unpaired Student’s t test. The data shown are representative of three independent experiments