Fig. 5.

Immunocytochemistry and western blotting of WT and R1441C LRRK2 MEF cells using various antibodies. A) Cells were incubated in the presence or absence of MLi-2 (200 nM, 2 h), and extracts subjected to western blotting using the indicated antibodies. B) Example of MEF cells stained with anti-LRRK2 antibody (N241A/34) and DAPI. C) Example of MEF cells stained with anti-LRRK2 antibody (UDD3) and DAPI. (D) Example of MEF cells in the absence or presence of MLi-2 (200 nM, 2 h) as indicated, and stained with knockout-validated RAB10 (SAB5300028) antibody and DAPI. E) Example of MEF cells in the absence or presence of MLi-2 and stained with an anti-pT73-RAB10 antibody (ab241060) and DAPI. F) Example of MEF cells in absence or presence of MLi-2 and stained with pS1292-LRRK2 antibody (Triton-X100/PBS-containing buffer conditions) and DAPI. G) As in (F), but cells stained with pS1292-LRRK2 antibody (PLA buffer conditions) and DAPI. Scale bars, 10 μm. Representative western and images are from two independent experiments with comparable results obtained in both cases.