Fig. 6: Functional heterogeneity of macrophage subsets and the contribution of monocyte-derived macrophages to L. infantum control.

a, b Relative expression of lipid and iron metabolism (a), and chemokines and cytokines (b) genes from sorted CD45.2⁺F4/80⁺CD11b^intCD64⁺ cells at 42 d.p.i. Data was downsampled to a maximum of 500 cells per cluster. c Number of iNOS⁺F4/80⁺ granulomas in WT mice, quantified from immunofluorescence images. Data pooled from 2 independent experiments (n = 7 for 19- and 42 d.p.i. and n = 6 for 72 d.p.i.). d Representative images showing F4/80(cyan), CLEC4F(white), TIM-4(red), and iNOS(green) in WT livers. Scale bars, 30 μm. e Number of F4/80⁺iNOS⁺ macrophages in WT livers at 42 d.p.i. Data pooled from 5 independent experiments (n = 19). f Representative images showing F4/8(cyan), CLEC4F(white), TIM-4(red), and iNOS(green) (top), F4/80⁺iNOS⁺ granulomas (bottom) in WT liver at 42 d.p.i. Scale bars, 100 μm. g Single-cell expression of Nos2, chemokines, and cytokines in CD45.2⁺F4/80⁺CD11b^intCD64⁺ cells at 42 d.p.i. h Frequency of infected cells within F4/80⁺ granulomas, obtained from 2-3 regions from each liver (ROIs=24 for 19 d.p.i., ROIs=23 for 42 d.p.i.). Data pooled from 2 independent experiments for each time point, 8 mice at 19- and 42 d.p.i., 7 mice at 72 d.p.i. i Representative images of F4/80⁺ macrophages in WT livers, stained with anti-F4/80(cyan), anti-CLEC4F(white), anti-TIM-4(red), and anti-L. infantum(yellow). Scale bars, 30 μm. j Number of F4/80⁺ granulomas quantified from immunofluorescence images. Data pooled from 2 independent experiments (n = 7 for 19- and 42 d.p.i., n = 6 for 72 d.p.i.). k Distribution of parasites inside and outside F4/80⁺ granulomas. Data pooled from 2 independent experiments (n = 7). l Parasite loads in WT and Ccr2^-/- mice at 42 d.p.i. Data pooled from 3 independent experiments (n = 12 for WT and n = 13 for Ccr2^−/−). m Number of L. infantum amastigotes per field in 42-day infected WT and Ccr2^−/− mice, quantified from confocal microscopy images. Data pooled from 2 independent experiments (n = 8). n Representative images showing F4/80(cyan), CLEC4F(white), TIM-4(red), and L. infantum(yellow) in WT and Ccr2 ^−/− livers at 42 d.p.i. Scale bars, 20 μm. Values in (c, e, h, j–m) represent the mean ± SD. P values were obtained in (c, e) using ordinary one-way ANOVA with Tukey’s multiple comparisons test, in (h, j) using Kruskal-Wallis test with Dunn’s multiple comparisons tests, in (k–m) using a two-tailed unpaired t test. Source data are provided as a Source Data file.