Extended Data Fig. 4: Phenotypical and functional effects of knockdown of microglial TGFα and VEGF-B. | Nature

Extended Data Fig. 4: Phenotypical and functional effects of knockdown of microglial TGFα and VEGF-B.

From: Microglial control of astrocytes in response to microbial metabolites

Extended Data Fig. 4

a, Quantification of astrocyte numbers in spinal cord sections of knockdown mice. SOX9-positive astrocytes per mm2 were quantified in spinal cord sections of four mice per group. b, IMARIS reconstruction of GFAP+ astrocytes in spinal cord sections as in a, and quantification of dendrite length, branches, volume, terminal points, and segments of n = 4 mice per group. c, d, qPCR analysis of Tgfa and Vegfb expression in sorted CNS-infiltrating inflammatory monocytes (c) and microglia (d) from mice injected with pCD11b-shControl, pCD11b-shTgfa, and pCD11b-shVegfb 7 days after EAE induction. Representative of two independent experiments with three biological replicates. e, qPCR analysis of Erbb1 and Flt1 expression in mice injected with pGFAP-shControl, pGFAP-shErbb1, and pCD11b-shFlt1 7 days after EAE induction. Representative of two independent experiments with three biological replicates. f, Left, flow cytometry analysis of VEGF-B and TGFα expression in microglia from mice injected with pCD11b-shControl, pCD11b-shTgfa, and pCD11b-shVegfb 7 days after EAE induction. Right, quantification of VEGF-B- and TGFα-positive microglia in n = 5 mice per group. Representative of two independent experiments with five biological replicates. g, Left, flow cytometry analysis of FLT-1 and ErbB1 expression in astrocytes from mice injected with pGFAP-shControl, pGFAP-shErbb1, and pCD11b-shFlt1 7 days after EAE induction. Right, quantification of FLT-1 and ErbB1-positive microglia in n = 5 mice per group. Representative of two independent experiments with five biological replicates. h, Naive mice were injected with lysolecithin, VEGF-B, or PBS into the corpus callosum by stereotaxic injection and 6 days later, brains were analysed by myelin staining. Representative of two independent experiments with five biological replicates. Data are mean ± s.e.m. P values were determined one-way ANOVA followed by Tukey’s post-hoc test (ah). 

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