Fig. 1: Spatially and temporally distinct repopulation kinetics of CAMs and MG.
a, Overview of microglia (MG) and subsets of CAMs during homeostasis in the CNS. dCAMs, dural CAMs; sdCAMs, subdural CAMs, including leptomeningeal (lmMΦ) and perivascular (pvMΦ) macrophages. b, Left: Scheme of the experimental layout. Right: CNS macrophage repopulation in complementary myeloid-specific fate mapping mouse lines was analyzed by confocal microscopy, flow cytometry, RNA-seq and scATAC-seq, followed by systemic LPS challenge and a middle-cerebral artery occlusion (MCAO) model of thromboembolic stroke. c,d, Representative flow cytometry plots of MG and sdCAMs after depletion (c) and quantification of their numbers per brain hemisphere (d). Symbols represent individual mice, n = 3 (CTL and 1 d BLZ945), n = 4 (8w, 12w, 26w BLZ945), n = 5 (5 d BLZ945), n = 6 (2w BLZ945), mean ± s.e.m. Ordinary one-way analysis of variance (ANOVA) with Šidák-adjusted multiple-comparisons P values. e, Representative confocal images from cerebral cortex, cerebellum and olfactory tubercle at 1 d, 5 d and 8w after BLZ945 treatment in WT mice. Arrows point to myeloid cells in their distinct compartments. f, Proportion of MG, lmMΦ and pvMΦ density at different time points and in diverse brain regions after BLZ945 treatment compared to controls (CTL). Symbols represent mean depletion at each time point, calculated as proportion of CTL mean. Error bars indicate s.e. of the depletion estimate, propagated from the standard errors of BLZ945 and CTL densities. n = 3 (CTL and 1 d BLZ945), n = 4 (8w, 12w, 26w BLZ945), n = 5 (5 d BLZ945), n = 6 (2w BLZ945), logistic growth modeling, t90%: time point in days when the repopulation reaches 90% compared to controls. R2: coefficient of determination for model equation and data points). Unpaired two-tailed Welch’s t-test with Bonferroni-corrected P values.
