Fig. 2

pcDNA3.1(+)-alpha-2 giardin and pcDNA3.1(+)-alpha-7.3 giardin activate mouse peritoneal macrophage NLRP3 inflammasome. The recombinant eukaryotic expression plasmids pcDNA3.1(+)-alpha-2 giardin and pcDNA3.1(+)-alpha-7.3 giardin were transfected into primary mouse peritoneal macrophages and cells or the supernatants were collected at 24 h for analysis of the expression, oligomerization, secretion and location of inflammasome key proteins. The PBS-only group (C) and pcDNA3.1(+) single treatment group were used as the negative controls, and the GEV treatment group was used as the positive group. a NLRP3 inflammasome key proteins, including NLRP3, pro-IL-1β, pro-caspase-1 and caspase-1 p20, were detected using western blotting. b IL-1β secretion levels in the supernatants were detected using an enzyme-linked immunosorbent assay (ELISA) assay. Differences between the control group and experimental groups were analyzed by one-way analysis of variance (ANOVA) using SPSS version 22.0 software. Asterisks indicate a significant difference between groups at **P < 0.01 and ***P < 0.001. c The ASC oligomerization levels were determined in the pellets using the DSS crosslinking assay, whereas the ASC levels in the cell lysates were used as a loading control. d ASC localization was observed using immunofluorescence. e NLRP3 localization was observed using immunofluorescence. ASC, Apoptosis speck-like protein; IL, interleukin; NLRP3, nucleotide-binding oligomerization-like receptor 3; n.s., not significant (P > 0.05)