Extended Data Fig. 2: Topical and molecular regulation of TGFα and VEGF-B. | Nature

Extended Data Fig. 2: Topical and molecular regulation of TGFα and VEGF-B.

From: Microglial control of astrocytes in response to microbial metabolites

Extended Data Fig. 2

a, Ingenuity pathway analysis of differentially regulated pathways in astrocytes from n = 3 control versus CX3CR1-AHR mice per group during EAE. b, Tgfa and Vegfb expression determined by qPCR in microglia from brain, cerebellum and spinal cord 21 days after EAE induction (left). n = 8 mice per group. c, Predicted NF-κB and AHR responsive sites (NREs and XREs, respectively) in Vegfb and Tgfa promoters. d, Microglia were isolated by FACS sorting from control and CX3CR1-AHR mice during EAE. Ex vivo ChIP assay of NF-κB p65 or AHR binding to predicted binding sites in the Vegfb promoter. n = 3 mice per group. Data are representative of two independent experiments. e, Reporter assay using a construct in which the Vegfb promoter controls luciferase expression (pVegfb-Luc). Luciferase activity was measured in HEK293 cells 24 h after transfection with pVegfb-Luc, pTK-Renilla, and plasmids expressing AHR or NF-κB p65. Data are representative of two independent experiments with four biological replicates. f, Ex vivo ChIP assay as in d for AHR binding to the Tgfa promoter. n = 3 mice per group. Representative of two independent experiments. g, Reporter assays using a construct in which the Tgfa promoter controls luciferase expression (pTgfa-Luc). Luciferase activity was measured in HEK293 cells 24 h after transfection with pTgfa-Luc, pTK-Renilla, and plasmids expressing AHR or control. Data are representative of two independent experiments with three biological replicates. Data in a, dg are mean ± s.e.m. P values were determined by one-way ANOVA followed by Tukey’s post-hoc test (b, d, e, f) or two-sided Student’s t-test (g). 

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