Journal: bioRxiv

Article Title: CANVAS causing AAGGG repeat expansions cause tissue-specific reduction in RFC1 expression and increase sensitivity to DNA damage

doi: 10.1101/2025.11.18.688292

Figure Lengend Snippet: A-C. Locomotor activity in adult male driver/shRNA alone controls flies, or flies expressing Gnf1 shRNA in post-mitotic neurons ( nsyb > gnf1 shRNA), of varying ages, quantified as beam breaks across a 24 h period in the Drosophila activity monitor (DAM). A : 3-7 days old flies (n=9-18); b : 21-23 days old flies (n=19-60); and C . 40-42 days old flies (n=15-23). D. Survival curve showing the percentage survival of nsyb > gnf1 shRNA flies compared to controls during normal aging. n=100 per genotype. E-G . DNA damage accumulation in control and neuronal Gnf1 knockdown adult male fly brains, quantified as the ratio of H2Av immuno-fluorescence to DAPI fluorescence at 12 days old ( e : n=5-7) and 40-42 days old ( f : n=9-11). G . Representative confocal images illustrating H2Av and DAPI staining in 40-42 days control and neuronal Gnf1 knockdown adult male fly brains. H2Av immuno-fluorescence is color-coded to indicate signal intensity. Scale: 100 µm. Error bars in A-C , E , F : standard error of the mean (SEM). Central line in dot plots: mean. P values are indicated, acquired via Brown-Forsythe and Welch ANOVA test with Dunnett’s T3 multiple comparisons post-hoc test (A,E,F) or Kruskal-Wallis test with Dunn’s T3 multiple comparisons post-hoc test (B, C) or Mantel-Cox Log rank test (D)

Article Snippet: Brains were incubated overnight at 4°C in a primary antibody against the phosphorylated form of H2Av (mouse UNC93-5.2.1, Developmental Studies Hybridoma Bank) at a final concentration of 1:200.

Techniques: Activity Assay, shRNA, Expressing, Control, Knockdown, Fluorescence, Staining