Fig. 4: Clonality and spatial distribution of macrophage subsets during late-stage VL.

a Intravital microscopy images representing a 42-day infected liver from a Clec4f^Cre-TdTConfetti mouse, stained with anti-CD31 (magenta), and showing tdTomato (red-nucleus), RFP (red-cytoplasm), YFP (yellow-cytoplasm), GFP (green-nucleus), and CFP (blue-membrane) KCs. Scale bars, 50 μm. b Frequency of KC clones in granulomas based on KC colors. Data pooled from live imaging performed in 2 independent experiments (n = 2 and quantification of 284 granulomas). c Representative rendered immunofluorescence image of a 42-day infected, wild-type liver showing the spatial distribution of CLEC4F⁺TIM-4⁺resKCs (yellow), CLEC4F^-TIM-4⁺KCs (red), CLEC4F⁺TIM-4^- moKCs (magenta), CLEC4F^-TIM-4^-momacs (cyan), and sinusoids (green). Scale bars, 30 μm. d Bar graphs showing the localization of each macrophage subset according to their interaction with the sinusoids in 42-day infected and naïve, wild-type mice. Data pooled from 2 independent experiments using 4 naïve mice and 8 infected mice (42 d.p.i.). Frequencies were calculated from four regions of interest (ROIs) per infected mouse (n = 32 ROIs) and 2-3 ROIs per uninfected mouse (n = 11 ROIs). e Scatter plots from immunofluorescence images showing the frequency of each F4/80⁺ population based on their distribution outside or inside granulomas at 42 d.p.i. Data pooled from 3 independent experiments (n = 11). In (d, e) 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. Values from (b, d, e) represent mean ± SD. Source data are provided as a Source Data file.