The percentage of Triton-X insoluble S129A aSyn increases during clearance.
(A) WT and S129A aSyn-GFP protein levels in yeast cells assessed by western blot analysis of total protein extracts at the indicated time points of aSyn-GFP clearance (left panel). Densitometric analysis of the immunodetection of aSyn relative to the intensity obtained for GAPDH, used as loading control, presented in arbitrary units (a.u.) (right panel) (***p<0.001, *p<0.05; one way ANOVA with Bonferroni's multiple comparison test). (B) Triton-X soluble (TS) and Triton-X insoluble (TI) fractions of aSyn WT or S129A assessed by western blot analysis (upper panel) and determination by densitometric analysis of the percentage of TI aSyn, at the indicated time points of aSyn clearance (lower panel) (*p<0.05; Mann-Whitney test). GAPDH was used as an internal control for the experiment. A representative result is shown from at least four independent experiments. Values represent the mean ± SD.