1. Pathogenic LRRK2 mutants co‐localize with Rab29 and disperse Golgi membranes
Pathogenic LRRK2 mutants co‐localize with Rab29 and disperse Golgi membranes HeLa cells were transfected with LRRK2‐G2019S or Myc‐Rab29 or LRRK2‐G2019S and 24 h later transfected with Myc‐Rab29. After 48 h, cells were permeabilized by liquid nitrogen freeze–thaw to deplete cytosol and then fixed and stained with mouse anti‐p115, mouse anti‐Myc, and rabbit anti‐GFP or rabbit anti‐LRRK2 antibodies. Myc‐Rab29 (green) and p115 (red) show co‐localization at the Golgi. Left and right panels were treated with or without MLI2 (200 nM, 4 h) as indicated.Myc‐Rab29 (red) and eGFP‐LRRK2‐G2019S (green) show co‐localization and dispersed Rab29‐labeled Golgi membranes.Cells treated with MLI‐2 (200 nM, 4 h) show compact, Rab29‐positive Golgi, and associated LRRK2‐G2019S (green).Percent of cells with compact Rab29 staining; ***P = 0.0002; **P = 0.002 with Student's unpaired, two‐tailed t‐test. Error bars represent SEM for three experiments with > 30 cells per condition in each experiment.LRRK2‐G2019S alone showing punctate staining throughout the cell.Data information: Scale bars, 10 μm.
Elena Purlyte et al. EMBO J. 2018;37:1-18
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