Fig. 2

Time-course of target inhibition and rephosphorylation of LRRK2 and Rab GTPase substrates across brain and peripheral tissues. G2019S LRRK2 knock-in mice were given a single 10 mg/kg MLi-2 dose via oral gavage and sacrificed at various time points. Brain, kidney, and lung homogenates were analyzed by western blot (n = 4 per group; 60 μg of protein were loaded per well) a. Maximal LRRK2 inhibition was achieved at 1 h post dose as shown by S1292 and S935 dephosphorylation while complete rephosphorylation was achieved by 24 h b, c. Rab10 was dephosphorylated by 1 h post-dosing in kidney and lung but not in the brain that showed variability in phosphorylated levels and did not respond to treatment a, d. Rab12 showed robust dephosphorylation by 1 h in brain and periphery a, e. Rab29 responded with maximal dephosphorylation in brain and lung by 1 h while kidney showed a slower but more significant response a, f. All three Rab GTPases recovered their phosphorylation levels by 12 h post-dose. These results demonstrate that Rab GTpases are dephosphorylated within the first hour following LRRK2 inhibition and rephosphorylate quicker than LRRK2 autophosphorylation recovery following wash-out conditions (~ 12 h vs ~ 24 h)