Fig. 4

Centrosome splitting in human dermal fibroblasts and lymphoblasts from G2019S mutant LRRK2 PD patients compared to healthy controls. a Example of control (ctrl) and G2019S mutant LRRK2 PD patient fibroblast stained with pericentrin antibody and DAPI. Scale bar, 10 μm. b Centrosome phenotype was quantified from 300 cells per line, and from 5 control and 5 G2019S mutant LRRK2 fibroblast lines. Control or G2019S mutant LRRK2 fibroblasts were treated with LRRK2-IN-1 (500 nM) or GSK2578215A (500 nM) for 60 min. Bars represent mean ± s.e.m. (between five independent lines). ***, p < 0.005; **, p < 0.01; *, p < 0.05. c Example of control (ctrl) and G2019S mutant LRRK2 PD patient lymphoblast stained with pericentrin antibody and DAPI. Scale bar, 10 μm. d Centrosome phenotype was quantified from 200 to 300 cells per line, and from three control and three G2019S mutant LRRK2 lymphoblast lines. Control or G2019S mutant LRRK2 lymphoblasts were treated with MLi2 (10 nM or 100 nM) or GSK2578215A (500 nM) for 2 h. Bars represent mean ± s.e.m. (between three independent lines). ***, p < 0.005. e Cells were either left untreated or incubated with 10 nM MLi2 as indicated, and extracts analyzed for phosphorylated (p-S935) or total LRRK2. f Quantification of S935 dephosphorylation in either control or G2019S-mutant LRRK2 lymphoblasts as indicated, in either the absence or presence of 10 nM MLi2. Bars represent mean ± s.e.m. (n = 3 lines each); ***, p < 0.005