Fig 1: Restoration of selected genes in SC of BAC-Q72 mice with ASO7 treatment. (A) Effect of ASO7 on Fyco1, C3 and Cyp51a1. Expression determined by qPCR relative to Gapdh. Reduced expression of C3 is significantly increased by ASO7, while increased expression of FycoI is significantly decreased by ASO7. Reduced Cyp51a1 expression was not improved by ASO7. (B) Effect of ASO7 on the expression of Eaat2 (Slc1a2), Pcp4, Ifih1, Trim30, p-Ampk, Sting, Cyp51a1, Tbk1, mTor, p62 and Lc3 determined by western blotting. (C) Densitometric quantifications of western blots (n = 3–5 mice). Bonferroni corrected Student’s t-tests. NS, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Fig 2: Validation of expression between BAC-Q72 mice and wild-type littermates for the top 2–4 hub genes identified in the lightgreen (A), midnightblue (B) and yellow (C) modules and selected other DEGs determined by qPCR. (D) Validation for Pcp4, an ALS-related gene, which was not assigned to a module by WGCNA. Values shown are means and SD. All individual wild-type versus BAC-Q72 comparisons were significant at P < 0.01. n = 5 and 3 mice for the wild-type and BAC-Q72 groups, respectively (A). n = 5 and 3 mice for all wild-type and BAC-Q72 groups, respectively, except for Ddx58 for which n = 6 mice per group (B). n = 6 mice per group (C and D). Probabilities were determined from unpaired two-tailed Student’s t-tests: **, P < 0.01; ***, P < 0.001.
Fig 3: Silencing of STAU1 mitigates SCA2 phenotypes. a Stau1 haploinsufficiency improves abnormal motor behavior of ATXN2Q127 mice as determined by rotarod behavior at 8, 12, 16, and 20 weeks of age. ATXN2Q127;Stau1+/- mice (green) have improved rotarod performance compared with ATXN2Q127 littermates (red) starting at 12 weeks of age. Note that Stau1 haploinsufficiency (orange) by itself does not alter motor function; n = 9–15 mice per group. Values shown are mean ± SE. Significance was determined using generalized estimating equations (GEE). NS, nonsignificant, *P < 0.05, **P < 0.01. b, c Reduction of Stau1 in vivo improves levels of key cerebellar proteins towards normalization. b Western blotting of cerebellar extracts from ATXN2Q127;Stau1+/- mice showing improvement of protein levels for Calb1, Pcp2, Rgs8, Pcp4, Homer3, and Fam107b towards normalization. Each lane represents cerebellar extract from an individual mouse. β-Actin is used as a loading control and the blots are from three replicate experiments. c Quantitative analysis of western blots shown in b. Data are mean ± SD, **P < 0.01, ***P < 0.001, Student t-test. d Combined immunostaining of ATXN2 (red) and Stau1 (green) of cerebellar sections from ATXN2Q127 and crossed ATXN2Q127;Stau1+/- mice (34 weeks of age) demonstrating reduced ATXN2-Stau1 aggregates in crossed ATXN2Q127;Stau1+/- mice. Scale bar, 30 µM. e Model for STAU1 in the pathology of SCA2 and other neurodegenerative diseases
Supplier Page from Abcam for Anti-PCP4 antibody