Fig. 2: TIM-4^- macrophages are monocyte-derived during late-stage VL.

a Representative contour plots showing the percentages of chimerism in F4/80⁺CD11b^intCD64⁺ cells from naïve and 42-day infected CD45.1⁺ parabiont. b Percentages of chimerism in F4/80⁺CD11b^intCD64⁺ cells from uninfected and infected CD45.1⁺ parabiotic partners at 42 d.p.i. c Frequency of CD45.2⁺ cells in infected CD45.1⁺ parabionts, gated on F4/80⁺CD11b^intCD64⁺CLEC4F^+/-TIM-4^+/- macrophages or Ly6C⁺CD11b⁺Ly6G^-SiglecF^- monocytes. Data pooled from two independent experiments (n = 3 for uninfected and n = 4 for infected pairs). d, e Number of monocytes in the livers of naïve (d) and 19- and 42-day infected WT and Ccr2^−/− mice (e) determined by flow cytometry. Data pooled from 5 independent experiments (naïve, n = 14 for WT and n = 17 for Ccr2^−/−; 19 d.p.i., n = 7 for WT and n = 9 for Ccr2^−/−; 42 d.p.i., n = 12 for WT and n = 14 for Ccr2^−/−). f Number of F4/80⁺ cells in WT and Ccr2^−/− at 42 d.p.i., quantified from confocal microscopy images. g Frequency of macrophage subsets based on CLEC4F and TIM-4 expression in WT and Ccr2^−/− mice at 42 d.p.i., quantified from confocal microscopy images. Data pooled from 2 independent experiments (n = 8 for WT and n = 9 for Ccr2^−/−). In (b–g) values show mean ± SD. In (d, f ) for data that passed the normality test, P values were obtained using a two-tailed unpaired t test. For data that did not pass the normality test, P values were obtained using a two-tailed Mann-Whitney test. In (c) P-values were obtained using ordinary one-way ANOVA with Tukey’s multiple comparisons, in (e) P-values were obtained using ordinary one-way ANOVA with Sidak’s multiple comparisons. h Representative dot plots from confocal microscopy images showing the frequency of macrophage subsets in infected WT and Ccr2^−/− mice at 42 d.p.i. Numbers indicate mean ± SD percentage of cells in the gate. i Representative rendered confocal microscopy images of 42 d.p.i. livers from WT and Ccr2^−/− mice showing CLEC4F⁺TIM-4⁺ (yellow) and CLEC4F^-TIM-4⁺ (red) macrophages, CLEC4F⁺TIM-4⁻ (magenta) moKCs and CLEC4F^-TIM-4^- (cyan) momacs. Scale bars, 200μm. Source data are provided as a Source Data file.