Extended Data Fig. 3: Generation and characterization of Il34-deficient and Csf1-deficient mice.
From: Negative feedback control of neuronal activity by microglia
a, Violin plots show the expression levels of cell-type specific representative marker genes across the 10 identified cell types from striatum snRNA-seq data analysis. Black dots indicate mean expression of selected gene per cell type. b, In situ hybridization for Il34 (left) and Csf1 (right) mRNAs show differential, region-specific expression in cortex, striatum, CA1, dentate gyrus (DG), CA3, corpus callosum (CC), and cerebellum of wild-type mice (WM: white matter, GM: grey matter, ML: molecular layer, GCL: granule cell layer, scale bar = 100μm). c, h, The striatal grey matter-specific or white matter-specific microglia depletion was achieved by breeding NestinCre/+ mice to Il34fl/fl mice or Csf1fl/fl mice, respectively, to generate Il34fl/fl; NestinCre/+ (purple, c) and Csf1fl/fl; NestinCre/+ mice (blue, h). d, i, Dot plots showing relative expression levels of Il34 and Csf1 mRNA normalized to Gapdh in the striatum of Il34fl/fl; NestinCre/+ mice (d) or Csf1fl/fl; NestinCre/+ mice (i) compared with littermate controls (d, n = 4 mice each, Il34 P < 0.0001, Csf1 P = 0.69; i, n = 3 and 5 mice, Il34 P = 0.07, Csf1 P < 0.0001, unpaired two-tailed t-test). e, Dot plots show the average microglia density per mm2 per mouse in cortex, striatum, cerebellum (cortex: n = 9, 12, and 10 mice, P < 0.0001, striatum: n = 9, 13, and 10 mice, P < 0.0001, cerebellum: n = 7, 7, and 8 mice, P = 0.34, One-way ANOVA with Tukey’s post hoc test). f, left, Dot plot shows levels of IL34 protein as determined by western blot analysis of striatal protein lysate from Il34fl/fl, Il34fl/+; NestinCre/+ or Il34fl/fl; NestinCre/+ mice normalized to DARPP32 expression (n = 3 mice, P = 0.0077, One-way ANOVA with Tukey’s post hoc test). g, j, Bar graphs show the average percentage of white matter regions in striatal images (0.5mm × 0.5mm) used to count WM and GM microglia in control and mutant mice for the data shown in Fig. 2c and e. (g, P = 0.99, n = 4 and 3 mice, unpaired two-tailed t-test; j, n = 4 and 2 mice). For gel source data, see Supplementary Fig. 1. Data shown as mean ± s.e.m.
