Fig. 9.

LRRK2 activity in human autopsy brain from control or PD patients. FFPE axial midbrain blocks containing substantia nigra at the level of the red nucleus from control (ctrl) and iPD patients were sectioned at 5 μm thickness. A) After deparaffinization and rehydration, antigen retrieval was achieved at high pressure in citrate buffer (pH 6.0). After 48 h incubation with primary antibodies, PLA amplification was carried out for 130 min. Representative images for PLA pair pS1292-LRRK2 (N241A/34) are shown for control and iPD samples (n = 4 for each), stained for tyrosine hydroxylase (TH) and DAPI. As a control, an iPD brain section was subjected to PLA reaction without the pS1292 antibody (–1292). Scale bar, 40 μm. B) Sections were deparaffinized, autofluorescence was quenched by Sudan black solution followed by antibody incubations and PLA reactions exactly according to detailed published protocols [23]. Representative images for PLA pair pS1292-LRRK2 (N241A/34) are shown for control and iPD samples (n = 3 for each), stained for tyrosine hydroxylase (TH) and DAPI. As a control, an iPD brain section was subjected to PLA reaction without one of the secondary antibody probes (-MINUS). Scale bar, 40 μm.