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Fig. 6 | Acta Neuropathologica Communications

Fig. 6

From: Neuropathology of RAN translation proteins in fragile X-associated tremor/ataxia syndrome

Fig. 6

FMRpolyA staining in control and FXTAS tissues. a Protein sequence of FMRpolyA epitope. b Western blot using FMRpolyA antibody with and without blocking peptide and GFP in HEK293 cells that were either mock transfected (lanes1), transfected with EGFP-N1 plasmid (lanes 2) and ATG FMRpolyA100 GFP (lanes 3). GAPDH used as a loading control. c Immunocytochemistry of Mock, EGFP-N1 and FMRpolyA100GFP transfected HEK cells using FMRpolyA antibody. Nuclei stained using DAPI. Scale bar is = 10 μm. d Representative brain images from FXTAS patients stained using FMRpolyA antibody with and without blocking peptide at 4x (top), 20x (middle) and 60x (bottom) magnification. Nuclei stained with hematoxylin. Scale bars are 500 μm, 100 μm and 20 μm respectively. e Staining of control and FXTAS brain tissue using pre-bleed serum from FMRpolyA antibody production. Nuclei stained with hematoxylin. Scale bar = 20 μm. f Representative images from control (left) and FXTAS (right) hippocampus (upper panels) and cortex (lower panels) stained with FMRpolyA antibody. Nuclei stained with hematoxylin. Inset- 60x magnification. Scale bar = 20 μm. g Quantification of F represented as percent neurons with aggregates. Data from hippocampus (top) and cortex (bottom). Results expressed as means ± SEM; Mann-Whitney U-test **** p < 0.0001. h Graph showing the percentage of cells with FMRpolyA positive aggregates that are neurons or glia in hippocampus (top) and cortex (bottom) from FXTAS tissue. i Graph comparing average staining intensity for FMRpolyA between control and FXTAS hippocampus (top) and cortex (bottom). Mann-Whitney U-test * p < 0.05, ns = not significant

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