Extended Data Fig. 3: Increased mVenus-CAAX-positive ectosomes from GBA1 and LRRK2 patient-derived dopaminergic neurons.
From: Glucosylceramide-induced ectosomes propagate pathogenic α-synuclein in Parkinson’s disease
a, iPS cell-derived dopaminergic neurons fixed and immunostained for β3-tubulin and tyrosine hydroxylase. b, Percentage of tyrosine hydroxylase positive dopaminergic neurons in GBA1 and LRRK2 mutant lines and their isogenic controls. n = 4 independent inductions; mean ± SEM. c, Live-cell maximum intensity projection of LRRK2-G2019S and isogenic corrected control (corr) iPS cell-derived dopaminergic neurons expressing mVenus-CAAX. Boxed areas highlighting ectosomes forming at the plasma membrane are magnified below. d, Number of mVenus-CAAX positive buds per cell area in LRRK2-G2019S and control iPS cell-derived dopaminergic neuron. n = 3 independent experiments; mean ± SEM; Two-tailed one sample t-test. e-f, Size distribution of attached mVenus-CAAX-positive vesicles in GBA1 and LRRK2 mutant lines. independent experiments: n = 3 for GBA1 (total 118 vesicles) and n = 4 for LRRK2 (total 239 vesicles); mean ± SEM. g-h Cytotoxicity assessment of mVenus-CAAX expression in GBA1 and LRRK2 mutant lines and their isogenic controls with MTT assay. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. Graphs are depicted as superplots where biological replicates are shown in large shapes, and where applicable technical replicates are shown as small shapes.
