Fig. 6
From: Parkinson disease-associated mutations in LRRK2 cause centrosomal defects via Rab8a phosphorylation
Pathogenic LRRK2 causes kinase-dependent pericentrosomal/centrosomal accumulation of endogenous phospho-Rab8a. a, b Cells were transfected with pathogenic LRRK2 or with kinase-dead pathogenic LRRK2 as indicated, and stained using an anti-phospho-T72-Rab8a antibody preabsorbed either with dephospho-peptide (p-Rab8a) or with phospho-peptide (p-Rab8a + pp), or with an anti-phospho-T72-Rab8a antibody preabsorbed with dephospho-peptide upon incubation of cells with 100 nM MLi2 for 60 min prior to immunocytochemistry as indicated. Scale bar, 5 μm. c Quantification of the percentage of non-transfected or transfected cells displaying phospho-Rab8a staining colocalizing with centrosomes within a 3 μm diameter circle in either the absence or presence of antibody preabsorption with peptides or pretreatment of cells with MLi2 as described above. Around 50 cells were quantified per condition per experiment. Bars represent mean ± s.e.m., (n = 3 independent experiments); ****, p < 0.001. d Quantification of the percentage of non-transfected or transfected cells displaying phospho-Rab8a staining colocalizing with centrosomes as described above. Around 50 cells were quantified per condition per experiment. Bars represent mean ± s.e.m., (n = 3 independent experiments); **, p < 0.01