Fig 5. GSDMD regulates the L. mexicana neutrophil niche during the first days of infection.

(A) C57BL/6 and Gsdmd^-/- mice were infected i.d with metacyclic dsRed⁺ L. mexicana promastigotes and the number and frequency of neutrophils in the bone marrow, blood, and at the infection site was determined by flow cytometry at the indicated timepoints. The flow cytometry gating strategy is shown. (B) The frequency of CD45⁺CD11b⁺Ly6G⁺ neutrophils at 24 hours p.i. in bone marrow, blood, and infected ear is represented (C) The number of CD45⁺CD11b⁺Ly6G⁺ neutrophils present at the infection site at 24, 48, and 72 hours p.i., as analyzed by flow cytometry and (D) the number of L. mexicana-DsRed⁺ CD45⁺CD11b⁺Ly6G⁺ C57BL/6 and Gsdmd^-/- infected neutrophils is given for each genotype and analyzed time point p.i. (E). The parasite burden in the infected C57BL/6 and Gsdmd^-/- ear was determined by limiting dilution (LDA) analysis 48 hours p.i. (F) C57BL/6 and Gsdmd^-/- mice were similarly infected with L. mexicana and 21 days p.i, the parasite burden assessed by LDA in infected ears. Data are shown as mean ± SD and are representative of n>2 experiments per time point, with n>4 mice/group. Mann-Whitney U-test *p<0.05, **p<0.01.