Fig. 5

Pathologic confirmation of receptor tyrosine-protein kinase ERBB4 downregulation in reactive astrocytes. a Feature plot of ERBB4 expression. b Violin plot of ERBB4 expression across seven brain cell types (differential gene expression of astrocytes vs rest using Wilcoxon rank-sum test with Benjamini–Hochberg correction for multiple comparisons, *****p < 0.00001, 2.28 log-e fold change). Ast astrocyte, Ex excitatory neuron, In inhibitory neuron, Oli oligodendrocyte, Opc oligodendrocyte progenitor cell, Mic microglia, End endothelia. c Plot of ERBB4 denoised, normalized, and natural log-transformed gene expression versus pseudotime with linear regression modeling (Bonferroni adjusted p-value). Colored by nuclei density at each coordinate. d Co-immunofluorescence images of AD (top) and normal (bottom) brain tissues stained for DAPI, SOX9, and ERBB4. Arrows point to DAPI⁺SOX9⁺ERBB4⁺ astrocyte nuclei; arrow heads point to DAPI⁺SOX9⁺ERBB4⁻ astrocyte nuclei. e Co-immunofluorescence images of AD (top) and normal (bottom) brain tissues stained for DAPI, VIM, and ERBB4. Arrows point to DAPI⁺VIM⁻ERBB4⁺ nuclei; arrow heads point to DAPI⁺VIM⁺ERBB4⁻ reactive astrocyte nuclei. All scale bars correspond to 5 um. f Proportions of ERBB4⁺ nuclei among total nuclei for 10 normal and 10 AD brains (Fisher’s exact test, ****p < 0.0001, odds ratio = 1.979). Aggregated count data from SOX9 + ERBB4 and VIM + ERBB4 co-immunofluorescence images. g Proportions of ERBB4⁺ nuclei among SOX9⁺ astrocyte nuclei for 10 normal and 10 AD brains (Fisher’s exact test, ***p = 0.0004, odds ratio = 2.472). h Proportions of SOX9⁺ nuclei among total nuclei for 10 normal and 10 AD brain co-stained for SOX9 and ERBB4 (Fisher’s exact test, p = 0.9422, odds ratio = 1.012). i Proportion of ERBB4⁺ nuclei among VIM⁺ reactive astrocytes for 10 normal and 10 AD brains. No reactive astrocytes were observed in images of normal brains. Error bars represents standard error of the mean (SEM)