Fig. 6: Effect of acute LRRK2 kinase inhibition on phospho-LRRK2 and phospho-Rab in urinary EVs of macaques.

Urine was collected from macaques after different treatments and urinary EVs were isolated as described in materials and methods. a Western blot detection of total LRRK2, pS935-LRRK2, total Rab10, pT73-Rab10, and TSG101 in urinary EVs of NHPs treated with PF360 or solvent (5 mg/kg p.o., 2 and 6 h) (quadruplicate measures per sample). Note that two different dilutions of the calibration standard were used and these are separated by a molecular weight marker containing a band that is detected by the anti-pS935-LRRK2 antibody. b Quantification of (a). The values of phosphorylation ‘rate’ are indicative of the proportion of protein that is phosphorylated at the given site, see materials and methods. Statistical differences were tested using a Kruskal-Wallis test followed by a Dunn’s post-hoc test using the vehicle group as control. Note that values for pS935-LRRK2 were below the linear range of quantification at the 2 h time point and statistical anlaysis was not possible. Error bars represent standard error of the mean (s.e.m.). *P < 0,05 (N = 3). MW molecular weight marker.