Fig. 6: Ku80 knockdown rescues neuronal viability and mitigates p53-mediated neurodegeneration in sALS and FTD3 models.
From: The Ku80-p53-SIRT1 axis in DNA damage response contributes to sporadic and familial ALS and FTD
a Time-course analysis of neuronal viability using LDH release assay in sALS iMNs. Neurons were differentiated from three distinct sALS iPSC lines and transduced with control shRNA (red), Ku80 shRNA-1 (aqua), or Ku80 shRNA-2 (blueberry). Data are presented as mean ± SEM, n = 3 independent differentiations. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. Immunoblot (b) analysis and quantification of total p53 (c) and CC3 (d) protein levels in 3-month-old sALS iMNs after neurotrophic factor withdrawal. Data are presented as mean ± SEM, n = 3 independent differentiation experiments, each data point is the average of 3 iPSC lines. Statistical significance was assessed by two-way ANOVA with Sidak’s multiple comparisons test. e Time-course analysis of neuronal viability using LDH release assay in iPSC-derived neurons carrying the heterozygous FTD3-causing CHMP2B G26403C mutation, transduced with control shRNA (red), Ku80 shRNA-1 (aqua), or Ku80 shRNA-2 (blueberry). Data are presented as mean ± SEM, n = 6 samples per group, generated from three independent differentiations of iPSCs derived from two subjects. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. Western blot analysis (f) and quantification of CC3 (g) and total p53 (h), and SIRT1 (i) protein levels in 2-month-old FTD3 cortical neurons after neurotrophic factor withdrawal. Data are presented as mean ± SEM from four biologically independent Western blot samples per group, obtained from two independent differentiations of isogenic iPSCs derived from two subjects. Statistical significance was assessed by two-way ANOVA followed by Sidak’s multiple comparison test. Western blot analysis (j) and quantification (k) of Ku80 protein levels in heads of 10-day-old flies expressing CHMP2BIn5 under the control of the GMR-Gal4 driver. Data are presented as mean ± SEM (n = 7). Statistical significance was assessed by two-tailed Student’s t test. Representative images (l) and quantification (m) of retinal degeneration in CHMP2BIn5-expressing flies co-expressing control RNAi (against the mCherry gene) or Ku80 RNAi lines. The total number of flies per genotype is indicated in each column. Statistical significance was assessed by chi-square test. Significance levels were defined as *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; ns not significant. The exact p values and all source data are provided as a Source Data file.
