Fig. 6

Pathologic confirmation of transcription factor NFIA downregulation in reactive astrocytes. a Feature plot of NFIA expression. b Violin plot of NFIA 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, 2.53 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 denoised, normalized, and natural log-transformed NFIA expression vs 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 NFIA. Arrows point to DAPI⁺SOX9⁺NFIA⁺ astrocyte nuclei; arrow heads point to DAPI⁺SOX9⁺NFIA⁻ astrocyte nuclei. e Co-immunofluorescence images of AD (top) and normal (bottom) brain tissues stained for DAPI, VIM, and NFIA. Arrows point to DAPI⁺VIM⁻NFIA⁺ nuclei; arrow heads point to DAPI⁺VIM⁺NFIA⁻ reactive astrocyte nuclei. All scale bars correspond to 5 um. f Proportions of NFIA⁺ nuclei among total nuclei for 10 normal and 10 AD brains (Fisher’s exact test, ****p < 0.0001, odds ratio = 2.739). Aggregated count data from SOX9 + NFIA and VIM + NFIA co-immunofluorescence images. g Proportions of NFIA⁺ nuclei among SOX9⁺ astrocyte nuclei for 10 normal and 10 AD brains (Fisher’s exact test, ****p < 0.0001, odds ratio = 9.910). h Proportions of SOX9⁺ nuclei among total nuclei for 10 normal and 10 AD brains co-stained for SOX9 and NFIA (Fisher’s exact test, p = 0.6395, odds ratio = 1.083). i Proportion of NFIA⁺ nuclei among VIM⁺ reactive astrocytes for 10 normal and 10 AD brains. No reactive astrocytes were observed in images of normal brains. Error bars represent standard error of the mean (SEM)