Fig. 3: Analysis of LRRK2 and Rab levels in urinary EVs of PD patients compared to healthy controls.

Urine samples of PD vs control groups were collected at Lille University Hospital. Genotype and diagnosis confirmed for all subjects. In total, this cohort included 77 PD patients (41 men and 33 women, with 7 patients carrying the G2019S mutation) as well as 53 control subjects (22 men and 31 women). An overview of the clinical characteristics of the different groups is given in Table 2. EVs were isolated from the urine samples and total levels and phosphorylation rates of LRRK2 and Rab8 were measured together with TSG101 levels via western blotting (quadruplicate measures per sample). Results were expressed relative to a calibrator and represented as a histogram. Values of phosphorylation ‘rate’ are indicative of the proportion of protein that is phosphorylated at the given site, see materials and methods. Shown here are values for total LRRK2 (a), total Rab8a (b), TSG101 (c), and phosphorylation rates for pS910-LRRK2 (d), pS935-LRRK2 (e), pS1292-LRRK2 (f) and pT72-Rab8a (g). Note that histograms each show first 2 bars corresponding to the values obtained from the groups of healthy controls and idiopathic PD patients, then a third bar with values of PD patients carrying the G2019S mutation, the last 4 bars correspond to the same values as the healthy controls vs iPD, now separated out per gender. Significant differences are tested using the two-tailed Mann–Whitney test. Note that the values of the LRRK2-G2019S group are shown for information, but are not included in the statistical analysis due to the smaller group size (N = 7). Error bars represent standard error of the mean (s.e.m.). *P < 0,05; **P < 0,01.