Fig. 4.

LRRK2 kinase activity following treatment with H₂O₂ in HEK293T cells. HEK293T cells were transiently transfected with either 3xFLAG WT or G2019S LRRK2 for 48 h before being incubated with increasing doses of H₂O₂ (125 μM, 250 μM, 500 μM and 1000 μM) or nigericin (2 μM) for 90 min at 37°C. A) Western blot analysis of RAB10 phosphorylation (pT73) and LRRK2 phosphorylation (pS1292 or pS935) in cells treated with H₂O₂ or nigericin. B) Densitometric quantification of pS1292-LRRK2 normalized to total FLAG-LRRK2. Bars represent mean±S.E.M. (n = 4 independent transfections). C) Densitometric quantification of pS935-LRRK2 normalized to total FLAG-LRRK2. Bars represent mean±S.E.M. (n = 4 independent transfections). D) Densitometric quantification of pT73-RAB10 normalized to total GFP-RAB10. Bars represent mean±S.E.M. (n = 4 independent transfections; ^****p < 0.001; ^**p < 0.01; Dunnett’s multiple comparison test).