Mitochondrially anchored Rab29-GTP recruits LRRK2. (A) First four bars: Pearson’s correlation coefficients calculated from three independent experiments with ≥40 cells per condition between LRRK2 and Rab29 in HeLa cells expressing GFP-R1441G LRRK2 or D2017A LRRK2 and either mito-Rab29 WT or mito-Rab29 D63A after 24 h of expression with or without MLi-2. MLi-2 treatment was 200 nM at the time of transfection (first four bars). P values were calculated from unpaired t tests; *, P = 0.03; ***, P = 0.0007 (two tailed). Second four bars: Pearson’s correlation coefficients calculated from three independent experiments with ≥20 cells each; total Myc-Rab10 and mito-R1441G LRRK2 or mito-D2017A LRRK2 (fifth and sixth bars); LRRK2 recruited to mitochondria by mito-Rab29 and pRab10 (seventh bar); PM-LRRK2 and pRab10 (eighth bar). Error bars represent SEM from three experiments. (B) Light microscopy of a HeLa cell transfected with GFP-R1441G (green) and mito-Rab29 D63A (cyan). Scale bar, 10 µm.