Extended Data Fig. 5: Effects of MC21 depletion.

a, Mice were immunized with MOG_35-55 and animals received at the peak of disease six injections of either 50 μg of isotype control antibody (rat IgG2b) or 50 μg purified anti-CCR2 (MC21). Shown is the mean clinical course ±SEM. N = 6-7 mice per group and asterisk indicates statistical significance with * p < 0,05 and ** p < 0,005; unpaired two-tailed T-test. Data are representative of one experiment with six mice. b, FACS analysis (left) and quantification (right; mean ± SD) of Ly6C⁺ MHCII⁺ (IA^b) monocytes in the blood of isotype or MC21 treated mice. N = 3 mice per group, asterisk indicates statistical significance with p < 0,05; unpaired two-tailed T-test. The experiment was repeated three times with similar results. c, Analysis and quantification of splenic immune cells in EAE mice that received two injections of 50 μg isotype control antibody or 50 μg purified anti-CCR2. Shown are % of the respective cell populations out of CD45⁺ cells (n = 4 animals per group; experiment was performed twice with similar results; mean ± SD; asterisk indicates statistical significance with p < 0,01; unpaired two-tailed T-test). T_regs were identified as CD4⁺FoxP3⁺. d, Analysis and quantification of blood immune cells in EAE mice that received two injections of 50μg isotype control antibody or 50μg purified anti-CCR2 (MC21). Shown are % of the respective cell populations out of CD45⁺ cells (n = 4 animals per group; experiment was performed twice with similar results; mean ± SD; asterisk indicates statistical significance with p < 0,05; unpaired two-tailed T-test). e, Repetition of Fig. 3 in an independent mouse facility and with purified MC21 antibody. Wt animals received either of 50μg isotype control antibody or 50μg purified anti-CCR2 at the peak of disease for two consecutive days. Shown are the EAE courses during the experiment (day 16 PI, mean score in each group: isotype: 2.7 ± 0.3 SEM; MC21 3.0 ± 0.3 SEM; asterisk indicates statistical significance with p < 0,01; unpaired two-tailed T-test;). f, Projection of CD44⁺Ly6G⁻CD11b⁺ non-neutrophilic, non-microglial cells from isotype- (left) and MC21-treated (right) animals on the metacell model from Fig. 1. g, Bar plots showing enrichment (log₂ fold change) of myeloid groups in MC21-treated mice compared to isotype controls. Error bars represent 95% confidence intervals. 3 mice were pooled for MARS-seq analysis depicted in f,g and n = 232 cells from isotype- and 147 cells from MC21-treated mice were analyzed in f, g.

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