Fig. 4

PRICKLE1-induced LRRK2 puncta are not in the endosomal compartment but rather in signalosome-like structures, together with DVL2 polymers. a-e Immunofluorescence analysis of the subcellular localization of transiently overexpressed LRRK2. a Areas double positive for LRRK2-PRICKLE1 are negative for the lysosomal marker LAMP1. Arrows point to the partial co-localization of LRRK2 with LAMP1 in the membrane. Arrowheads point to LAMP1, which does not localize with LRRK2-PRICKLE1 puncta. b A late endosomal marker, RAB7, co-localizes with PRICKLE1 (arrowheads), but not with LRRK2 puncta (arrows). c LRRK2 forms puncta with DVL2. d PRICKLE1 and DVL2 are found in close proximity, but do not co-localize. PRICKLE1 is surrounded by 1 to 3 DLV2 complexes. e In the presence of LRRK2, DVL2 and PRICKLE1 partially co-localize (arrows). The co-localization of LRRK2, DVL2 and PRICKLE1 reveals the capacity of these proteins to form signalosome-like structures. Arrowheads show the localization of PRICKLE1 in close proximity to DVL2 puncta and LRRK2. Nuclear staining by Dapi is in blue. N ≥ 3. Scale bars show 10 μm