Efferocytosis of PIT and entrapped bacteria in vivo. Spleens from mice infected i.p. with control or FliC^ind GFP–S. Typhimurium for 24 h and treated with doxycycline for 3.5 h, were harvested, prepared for flow cytometry, and stained. Neutrophils with intracellular macrophage (MΦ) markers (CD45⁺ CD11b⁺ Ly6G^+high CD68⁺ F4/80⁺; A, B, and D), and neutrophils with intracellular MΦ markers and GFP–S. Typhimurium (CD45⁺ CD11b⁺ Ly6G^+high CD68⁺ F4/80⁺ GFP⁺; E) were identified by flow cytometry on an LSR II. (C, F, and G) Neutrophils were isolated by purification of Ly6G⁺ cells from spleen, stained, and analyzed by Amnis ImageStream. Experimental data from WT (six mice per group) and Ncf1^−/− mice (seven per control group and six per FliC^ind group) was pooled from two individual experiments. Data from six Casp1^−/−Casp11^−/− mice is representative of two individual experiments. The numbers of mice in all groups are noted in figure. Five Casp1^−/−Casp11^−/− mice were used per group and six animals per group were used for all other experiments. Control and FliC^ind GFP–S. Typhimurium strains are flagellin (flgB) mutants. Gating was performed using FlowJo (A, B, D, and E) or IDEAS (C, F, and G). P-values were determined by a two-tailed Student’s t test; *, P < 0.05.