Fig. 3
From: Parkinson disease-associated mutations in LRRK2 cause centrosomal defects via Rab8a phosphorylation
Pathogenic LRRK2 causes deficits in centrosome cohesion in SH-SY5Y cells. a Example of non-differentiated SH-SY5Y cells stably expressing GFP, or flag-tagged wildtype or G2019S-mutant LRRK2 as indicated, and stained for pericentrin and DAPI. Scale bar, 10 μm. b Quantification of the split centrosome phenotype in cells expressing GFP, or flag-tagged wildtype or G2019S-mutant LRRK2 as indicated. Around 20 cells with duplicated centrosomes were analyzed per condition. Bars represent mean ± s.e.m. (n = 4 independent experiments); *, p < 0.05. c Quantification of the split centrosome phenotype in cells expressing flag-tagged wildtype or G2019S-mutant LRRK2 as indicated, in either the absence or presence of kinase inhibitors (500 nM LRRK2-IN-1 or GSK2578215A for 1 h) as indicated. Around 20 cells with duplicated centrosomes were analyzed per condition. Bars represent mean ± s.e.m. (n = 3 independent experiments); **, p < 0.01; *, p < 0.05. d Cells were either left untreated or incubated with 500 nM GSK2578215A for 1 h, and extracts analyzed for phosphorylated (p-S935) or total (flag) LRRK2. e Quantification of S935 dephosphorylation in cells expressing flag-tagged wildtype or G2019S-mutant LRRK2 as indicated, in either the absence or presence of 500 nM GSK2578215A for 1 h. Bars represent mean ± s.e.m. (n = 3); *, p < 0.05