Fig. 2
LRRK2 mutations increase the extracellular ratio of pT73-Rab10 to total Rab10 as measured with single-molecule array assays (SiMOA) in serum. (A) Levels of total LRRK2, (B) total Rab10 and (C) the ratio of pT73-Rab10 to total Rab10 as measured by SiMOA in venous mouse serum from Lrrk2−/− (8 males and 3 females), non-transgenic C57BL6/J (12 males and 8 females), and knockin Lrrk2R1441C/R1441C mice (9 males and 8 females). (D-F) A cohort of transgenic Tg-WT-BAC Lrrk2 (4 males and 9 females) and Tg-G2019S-BAC Lrrk2 mice (5 males and 3 females) were also analyzed for these markers. In these strains, no differences were noted between males and females. Ages in both cohorts of the mice ranged from 3–6 months. (G) Representative immunoblots of lysates from HEK-293T cells transfected with FLAG-LRRK2 plasmids 24-hours before harvesting lysates. (H) Quantification of endogenous pT73-Rab10 to total Rab10 ratio. (I) SiMOA assay analysis of the lysates for the ratio of pT73-Rab10 to total Rab10. Dots show mean values calculated from independent experiments. Columns show group means, error bars show SEM, and p values are determined with two-tailed t-tests, or from one-way ANOVA followed by Tukey’s post-hoc test in the graphs with more than two groups