Figure 1.

Effect of LRRK2 on mitochondrial morphology. (A) Representative immunoblot shows LRRK2 levels in human dopaminergic neuroblastoma SH-SY5Y clonal lines that stably overexpress WT or mutant LRRK2. An aliquot of 20 μg/lane was loaded and tubulin used as an internal loading control. (B) SH-SY5Y cells were transfected with mito-DsRed2 to label mitochondria, fixed 2 days after transfection, immunostained with tubulin and evaluated. Representative pictures of positively transfected cells are shown. Green, tubulin; red, mito-DsRed2; blue, DAPi. Insets show enlargements of boxed areas. (C and D) Quantification of mitochondrial morphology revealed a significant decrease in the aspect ratio (C), and increase in the percentage of cells displaying fragmented mitochondria (D) compared with controls. For each cell line, at least 500 cells were analyzed in each experiment and experiments were repeated three times (*P < 0.001, Student's t-test).