Fig. 3

Substitution of Y133 and Y136 with alanine in synT blocks the aggregation-inhibiting effect of Fasudil. a H4 neuroglioma cells were treated with 20 μM Fasudil or vehicle and transfected with wt synT or the indicated mutants together with SYPH1. 24 h post-transfection, cells were fixed and immunostained against synT and SYPH1. Representative images show the expression of synT, SYPH1 and DAPI positive nuclei in the absence of Fasudil. Scale bar 25 μm. b Transfected cells with and without inclusions from 9 images per experimental conditions of n = 4 independent experiments were counted. Data are given as mean ± S.E.M, **p < 0.01, n.s. = not significant, T-Test. c Western blot analysis of H4 protein lysates from wt or indicated mutants 24 h after transfection revealed no significant changes in synT protein expression