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Fig. 1.
Neuronal-like SHSY5Y cells develop EGFP ataxin-3 protein aggregates. (A) SHSY5Y cells transiently expressing EGFP-fused human ataxin-3 (28Q and 84Q) develop protein aggregates. White arrowheads indicate EGFP-fused ataxin-3 protein aggregates. (B) The mean number of aggregates per transfected cell was calculated by averaging six experimental replicates (different fields of view) per coverslip. Aggregates were more commonly observed in cells transfected with the EGFP ataxin-3 84Q repeat expansion than cells transfected with EGFP ataxin-3 28Q repeat expansion. (C) Quantification of the percentage of cells harbouring protein aggregates revealed significantly more cells were affected by protein aggregates following transient transfection with EGFP ataxin-3 84Q compared to EGFP ataxin-3 28Q or EGFP only. (D) Western blotting was used to examine the expression of ataxin-3 following transient transfection of EGFP constructs. (E) Expression of EGFP-fused ataxin-3 was significantly higher in cells transfected with EGFP only or EGFP ataxin-3 84Q. Data are mean±s.e.m. n=4-5 experimental replicates. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001; ns, not significant (one-way ANOVA with Tukey's post-hoc test).