Fig. 2: Gsdmd disruption reduces, while Casp-1/11 disruption completely abolishes, IL-1β secretion.

a C. albicans-triggered GSDMD cleavage and IL-1β production in Casp-1/11^-/- mice. BMDMs from WT and Casp-1/11^-/- mice were infected with C. albicans for 6 h at MOI 50. GSDMD (cleaved p30 and full-length p53), pro-IL-1β, and mature IL-1β (m-IL-1β) in the cell lysates and m-IL-1β in the supernatants were assessed by western blotting. Results are representative of data from at least three biological replicates. The intensities of protein bands were quantified using ImageJ. Comparison was made by two-way ANOVA followed by Sidak’ multiple comparisons test. Data shown are means ± SD (n = 3 biologically independent samples). p = 0.003 (cell lysate-m-IL-1β), p < 0.0001 (supernatant-m-IL-1β) vs. WT. Source data are provided as a Source Data file. b IL-1β secretion by BMDMs from WT and Casp-1/11^-/- mice. BMDMs were infected with C. albicans (MOI = 10) for the indicated time periods. IL-1β in culture supernatants was measured by ELISA. Comparison was made by two-way ANOVA followed by Sidak’ multiple comparisons test. Data shown are means ± SD (n = 3 mice per group). p < 0.0001 (12 h, 16 h) vs. WT. c C. albicans-triggered IL-1β production in WT and Gsdmd^-/- mice assessed by western blotting. Comparison was made by two-way ANOVA followed by Sidak’ multiple comparisons test. Data shown are means ± SD (n = 3). p < 0.0001 vs. WT. Source data are provided as a Source Data file. d IL-1β secretion by BMDMs from WT and Gsdmd^-/- mice measured by ELISA. Comparison was made by two-way ANOVA followed by Sidak’ multiple comparisons test. Data shown are means ± SD (n = 3 mice per group). p = 0.0003 (4 h), p < 0.0001 (8 h, 12 h, 16 h) vs. WT. e Gsdmd disruption did not alter the total amount of m-IL-1β produced by C. albicans-challenged BMDMs. BMDMs from WT and Gsdmd^-/- mice were infected with C. albicans for 12 h at the indicated MOIs. The amounts of m-IL-1β in culture supernatants and cell lysates (intracellular) were measured by ELISA. Comparison was made by two-way ANOVA followed by Sidak’ multiple comparisons test. Data shown are means ± SD (n = 3 mice per group). p < 0.0001 vs. WT. f C. albicans-induced IL-1β production in Casp-1/11^-/- mice. WT and Casp-1/11^-/- mice were intravenously challenged with 1 × 10⁶ CFU C. albicans, and the level of IL-1β in the serum was assessed at the indicated time points. Two-tailed unpaired Student’s t test was used for statistical comparisons. Data shown are means ± SD (n = 4 mice per group). p = 0.0022 (8 h), p < 0.0001 (16 h), p = 0.0029 (24 h) vs. WT. g C. albicans-induced IL-1β production in Gsdmd^-/- mice. WT and Gsdmd^-/- mice were intravenously challenged with 1 × 10⁶ CFU C. albicans. Two-tailed unpaired Student’s t test was used for statistical comparisons. Data shown are means ± SD (n = 4 mice per group). p = 0.1542 (8 h), p = 0.0881 (16 h), p = 0.0003 (24 h) vs. WT. h Fungal burden in the kidneys, livers, spleens, and lungs of untreated or anakinra-treated WT and Gsdmd^-/- mice. Mice were sacrificed 2 days after intravenous injection of 1 × 10⁶ CFU C. albicans. Two-tailed unpaired Student’s t test was used for statistical comparisons. Data presented are the means ± SD (n = 7 for Control, n = 5 for Anakinra-treated mice) of three independent experiments. p = 0.0014 (kidney without anakinra), p = 0.8061 (kidney with anakinra) vs. WT. (i) Clinical score of C. albicans-challenged untreated or anakinra-treated WT and Gsdmd^-/- mice. Two-tailed unpaired Student’s t test was used for statistical comparisons. Data presented are the means ± SD (n = 7 for Control, n = 10 for Anakinra-treated mice) of three independent experiments. p = 0.0009 (12 h without anakinra), p = 0.0551 (12 h with anakinra), p < 0.0001 (24 h without anakinra), p = 0.5316 (24 h with anakinra) vs. WT. j Kaplan-Meier survival plots of C. albicans-challenged untreated or anakinra-treated WT and Gsdmd^-/- mice. Mice were intravenously challenged with 2 × 10⁵ CFU C. albicans and monitored for 15 days. Log-rank analysis was used. p = 0.0031 (without anakinra), p = 0.4337 (with anakinra) vs. WT.