Fig. 2

NEFH mutations cause protein aggregation in SH-EP. a Amino acid alignment of normal and mutant NEFH C-terminal parts reported in the CMT2cc presenting the Cryptic Amyloidogenic Element translation and their differences. Amino acid color correlates with polarity: hydrophobic in black, hydrophilic in green, acidic in red and basic in blue. b Confocal images of SH-EP transfected with eGFP tagged NEFH vectors and counterstained for tubulin in red and nucleus (DAPI) in blue. Scale bar: 20 μm. c Quantification of SH-EP cells with protein aggregates. Values represent mean +/− standard deviation of 15 fields, repeated in triplicates and analyzed by one-way ANOVA on ranks followed by Dunn posthoc test (P < 0.001). d 0.1% Triton X-100 Soluble and Insoluble fractionation revealed by western-blotting using GFP antibody to stain NEFH fusion protein and GAPDH gene to normalize