Extended Data Fig. 5: Effects of acivicin on microglia, macrophage and whole blood cell numbers.

a, Representative images showing morphology and density of primary microglia treated with vehicle or acivicin for 48 h stained with nuclei marker Hoechst or CellMask Red Whole Cell Stain. Quantification of the total number of microglia. Data are representative of one experiment with technical replicates (open symbols). Scale bar, 100 μm. b, Representative confocal images showing morphology and density of vehicle- and acivicin-treated BMDMs treated for 24 h. Scale bar, 100 μm. Insets depict digitally zoomed in regions of interest. Quantification of the total number of BMDMs in the field of view. Data are from n = 7 independent experiment. c, Representative flow cytometric plots of gating strategy for BMDMs treated with acivicin for 24 h or left untreated. Cell viability of single CD45⁺ cells was assessed by Sytox blue live/dead nuclei stain. Quantification of percent cell viability of BMDMs treated with acivicin or left untreated. Data are from n = 2 independent experiments with n = 4 mice per group. d, Quantification of complete blood count analysis of white blood cells (WBCs), red blood cells (RBC), platelets (PLT) (left) and composition of WBCs (right) of blood samples from healthy mice treated with saline or acivicin for 10 days. Data are from n = 8 mice (saline) or n = 7 mice (acivicin). e, Representative flow cytometric plots of cells expressing CD45 and CD11b from the spinal cord of healthy mice treated with acivicin or saline every day for 14 days. Quantification of total CD45^loCD11b⁺ microglia cell numbers in healthy mice treated with acivicin or saline. Data are from n = 3 mice per group. Each symbol represents an individual experiment (b) or mouse (c-e). Data are shown as means ± s.e.m. (b, d, e) or means ± s.d. (c). P = 0.68 or P > 0.99; n.s., not significant, as determined by two-tailed unpaired Student’s t-test (b) or two-tailed Mann-Whitney test (e).

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