Wild-type (WT) or gasdermin D knockout (Gsdmd−/−) mice were treated with 20 mg kg−1 of LPS i.p. or saline. After 6 h, plasma and spleens were collected for analysis. a Spleen lysates were immunobloted for NLRP3 and cleaved caspase-1 (p10) to assess for inflammasome activation, and quantified in b. Data indicate WT saline (n = 2), WT LPS (n = 3), Gsdmd−/− saline (n = 3), Gsdmd−/− LPS (n = 3) animals per group. NLRP3 upregulation and caspase-1 cleavage were similar in both LPS-treated WT and Gsdmd−/− mice. Plasma from each mouse was collected and analyzed for c IL-1β and d HMGB1 by ELISA. For c and d, data indicate WT saline (n = 13), WT LPS (n = 8), Gsdmd−/− saline (n = 9), Gsdmd−/− LPS (n = 5) mice per group. Note that Gsdmd−/− mice do not secrete IL-1β in response to LPS, but HMGB1 secretion is unchanged. Data with error bars are represented as mean ± SEM. Adjusted p values, provided in the panels, and n.s. not significant as determined by two-way ANOVA with Tukey’s multiple comparison correction, two-sided.