FIGURE 6.

Quantification of co‐localisation and overall expression of endogenous FUS, PA2G4 and TRA2β in CGG‐repeat RNA and FMRpolyG transfected HGrC1 cells. (A) HGrC1 cells were transfected with an empty plasmid or left untreated, and immunocytochemistry was used at 48 h post transfection to examine the cellular localisation of candidate proteins. Scale bars represent 20 μM. (B) HGrC1 cells were co‐transfected with a plasmid expressing 60x CGG repeats and RNA FISH followed by immunocytochemistry were used after 48 h to identify the colocalisation of CGG RNA aggregates and candidate proteins. Scale bars represent 10 μM. (C) Quantification of colocalisation from 40 individual cells over three separate experiments. Data are presented as the mean ± SEM. (D) Western blot for candidate protein expression following transfection of empty, Δ5′UTR FMR1 (CGG)100x GFP or 5′UTR FMR1 (CGG)100x GFP plasmids. Quantification of signal intensity is normalized to that of loading control ACTB or TUBA. Data are presented as the mean ± SEM of four individual experiments, Mann–Whitney test, *p < .05.