Fig. 5: GSDMD is important for the restriction of L. mexicana, L. major, and L. braziliensis in macrophages.

a Bone marrow-derived macrophages (BMDMs) from C57BL/6 (WT) and Gsdmd^–/– mice were pretreated for 4 h with LPS (100 ng/mL) and infected with stationary-phase L. mexicana, L. major, L. braziliensis MOI 10 for 24 h. IL-1β released in BMDM supernatant was measured by ELISA. b–n BMDMs from WT and Gsdmd^–/– mice were infected with stationary-phase parasites at an MOI 5 for 2 h, washed, and incubated for 1, 24, 48, and 72 h. Parasites used were: L. mexicana constitutively expressing GFP (b–f), L. major constitutively expressing dsRED (g–k), and L. braziliensis constitutively expressing GFP (l–p). b, g, l The percentage of infected cells was estimated by flow cytometry. e, j, o The mean fluorescence intensity of intracellular parasites was estimated by flow cytometry. c, h, m The percentage of infected cells was scored in Giemsa-stained cells. f, k, p The average number of amastigotes per cell was scored in the Giemsa-stained cells. A total of 100 cells in each triplicate well were analyzed. d, i, n Representative images of Giemsa-stained cultures showing intracellular amastigotes. Scale bar 50 µm. Data are presented as mean values ± SD of triplicate wells. *P < 0.05 comparing the indicated groups, as determined by two-way ANOVA. Shown is one representative experiment of three independent experiments performed. Source data are provided as a Source Data file.