Journal: Nature neuroscience
Article Title: C9ORF72-ALS/FTD-associated poly(GR) binds Atp5a1 and compromises mitochondrial function in vivo
doi: 10.1038/s41593-019-0397-0
Figure Lengend Snippet: a, Schematic of CamKII-tTA and (GR)80 constructs. b, Relative expression levels of (GR)80 mRNA in the frontal cortex of different CamKII;(GR)80 mouse lines at 2 months of age. Line 8 = 37.4±6.27, line 16 = 4.27±0.89, line 18 = 1.00±0, n = 2 mice per line. c, Age-dependent accumulation of poly(GR) in frontal cortex neurons of line 16 CamKII;(GR)80 mice (from 3 independently repeated experiments). Scale bar, 50 μm. d, Standard curve for the mesoscale discovery immunoassay with a synthetic (GR)8 peptide. Values are mean±s.d. of two independent experiments. e, Poly(GR) levels in frontal cortex tissues of CamKII;(GR)80 mice were measured by immunoassay; 3 months = 0.44±0.04, 7 months = 1.05±0.06, 12 months = 2.22±0.64 (n = 4 mice). Values are mean±s.e.m., F(2,9) = 86.57, **P = 0.0044, ****P= 0.0001 by one-way ANOVA with Tukey’s post hoc analysis for multiple comparisons. f, Poly(GR) levels in frontal cortex tissue of 2 control brains and 3 C9ORF72-related ALS/FTD brains measured by immunoassay. Control brain sample values were subtracted. Control brains = 0± 0, C9ORF72-related ALS/FTD brains = 7.34± 2.29. g, Neuronal types with different poly(GR) expression patterns in CamKII;(GR)80 mice at 8 months of age (repeated 3 times independently with similar results). Scale bar, 5 μm. h, Pie chart showing percentage of neuronal types A-D. Over 150 cells expressing poly(GR) were quantified. ECL, electrochemiluminescence.
Article Snippet: The immunoprecipitation samples were analyzed by western blot (as described above) with the following primary antibodies: anti-total OXPHOS antibody cocktail (Abcam, ab110413; 1:1,000), rabbit polyclonal anti-(GR)8 antibody (custom made by Covance, 1:100) and mouse anti-ATP5A1 (Abcam, ab14748; 1:1,000).
Techniques: Construct, Expressing, Electrochemiluminescence
