Extended Data Fig. 6: Microglial and neuronal Hexb jointly drive Sandhoff disease pathogenesis.
From: Microglia–neuron crosstalk through Hex–GM2–MGL2 maintains brain homeostasis
A) Schematic overview of genetic targeting. Hexbfl/fl were obtained by crossing the B6.Hexbtm1a(EUCOMM)Hmgu/H line with a FLP deleter. Newly generated Hexbfl/fl mice with LoxP sites (black triangles) around exon 2 were crossed to Cx3cr1Cre/+ and NesCre/+ mice. B) B)-C) Relative Hexb gene expression in FACS-sorted microglia (B) and bead-purified neurons (C) among different genotypes measured by qPCR. D) Relative Syt1 (neuronal marker gene), Gfap (astocytic marker gene), Itgam (microglia marker gene), and Plp1 (oligondendroglial marker gene) gene expression in bead-purified neurons among different genotypes measured by qPCR. E-F) Development of bodyweight (E) and muscle strength (F) for Hexbfl/fl (n = 15), Cx3cr1Cre/+:Hexbfl/fl (n = 15), NesCre/+:Hexbfl/fl (n = 15), and Cx3cr1Cre/+:NesCre/+:Hexbfl/fl (n = 14) mice. G) Representative immunohistochemical images of brain sections from the indicated genotypes. Top row: Microglia immunostained for P2RY12 (red), HEXB (brown), and counterstained with hematoxylin (Htx, blue). Bottom row: Neurons stained for NeuN (red), HEXB (brown), and Htx (blue). Insets show higher magnification views with yellow lines indicating the paths used for intensity profile quantification below. Triangles highlight intracellular HEXB-double positive structures. Graphs display greyscale intensity profiles of the deconvoluted staining signals along the yellow lines (red = P2RY12 or NeuN, brown = HEXB, blue = Htx). Data shown as mean ± s.e.m. Statistical analyses: one-way ANOVA with Tukey’s post hoc test (E-F). Illustrations in a were created using BioRender (https://biorender.com).
