Fig. 7: Astrocytic Nrf2 reduces Aß pathology.
A Schematic illustrating the crossing of APP/PS1 and GFAP-Nrf2 mice. B–D Brains were fixed in 4% of paraformaldehyde and embedded in paraffin. The brains were cut into 10 μm coronal sections. Antigen retrieval was performed using either formic acid or sodium citrate buffer prior to immunohistochemical staining with 6E10 antibody and processed by VECTASTAIN® Elite® ABC kit. (B) shows an example picture at low magnification, with (C) and (D) focusing on the cortex and hippocampus (CA1), respectively. Arrowheads indicate 6E10 staining in plaques and neurons of APP/PS1 and APP/PS1_X_GFAP-Nrf2 mice. Quantification of total plaque number (left), average diameter (middle) and percent area covered (right) in the cortex (E) and hippocampus (F). Mean ± SEM shown here and throughout this figure. *p = 0.0003, 0.0033 (E); 0.0015, 0.0047 (F), unpaired two-sided t-test (n = 4 per mice per genotype). G, H The level of human Aß42 (E) and Aß40 (F) in Triton-X, SDS, and urea fractions in cortex and hippocampus was quantified by sandwich ELISA. Two-way ANOVA (main genotype effect) for Aβ42: F (1, 18) = 6127, p < 2.9E−24 (cortex); F (1, 18) = 2357, p < 1.5E−20 (hippocampus). *p values for (G): 7.1E−09, 6.8E−09, 9.7E−17, 0.003, 4.2E−12 (Bonferroni post-hoc test). Two-way ANOVA (main genotype effect) for Aβ40: F (1, 18) = 161.2, p = 4.9E−17 (cortex); F (1, 18) = 161.2, p = 2.0E−10 (hippocampus). *p values for (H): 1.2E−09, 1.4E−15, 2.1E−25, 1.9E−07, 1.5E−06, 1.20E−22 (Bonferroni post-hoc test). N = 4 animals per genotype. I Western analysis of whole cortical extracts for levels of full-length APP, using the 22C11 antibody. One-way ANOVA F (3, 8) = 99.92, p = 1.1E−06 followed by Bonferroni’s post-hoc test (n = 3 mice per genotype). *p < 5.6E−05, 3.3E−06, compared to WT. J RNA-seq reads from both cortex and hippocampus spanning the KM/NL Swedish locus across all four genotypes were scored as WT or mutant, and the % calculated (n = 3 mice per genotype).
