Fig. 5

α-Synuclein interacts with LRRK2 to regulate autophagy. a. Total lgg-1::mCherry fluorescence in four CEP neurons and nerve ring from 5 day (adult day 3) nematode lines expressing α-synuclein and LRRK2 (WT, G2019S or Δlrk-1). b. Representative pictures of dat- 1::lgg-1::mCherry fluorescence in C. elegans lines expressing α-synuclein and LRRK2 (WT, G2019S or Δlrk-1). Scale bar represents 60 μm. c. Immunoblot of lgg-1::mCherry in lines of C. elegans lines expressing α-synuclein. Age-synchronized nematodes (50 nematodes/lane, adult days 1 and 3) were used. Actin is shown below to indicate relative protein load per well. d. A control experiment showing no change in levels of dat-1::GFP fluorescence in C. elegans lines expressing α-synuclein and LRRK2. Total fluorescence from all four CEP neurons from day 3 adult nematodes were plotted to detect any nonspecific general effects from α-synuclein and LRRK2 expression. e. Levels of lgg-1::mCherry fluorescence over the lifespan in C. elegans lines expressing α-synuclein ± LRRK2 (WT, G2019S or Δlrk-1). Total fluorescence of dat- 1::lgg-1::mCherry from CEP neurons and nerve ring was measured during a 14 day period using age synchronized nematodes. Total dat- 1::lgg-1::mCherry fluorescence was normalized to dat-1::GFP fluorescence in lines expressing α-synuclein ± LRRK2 (WT, G2019S or Δlrk-1), shown in panel D. The scales for the experiments in Figs. 3D and 5E are not meant to be quantitatively identical because the experiments were performed at different times. N = 40 animals/condition, **P < 0.001 and *P < 0.05 compared to lgg-1::mCherry. The G2019S LRRK2/α-synuclein line also differed from the WT LRRK2/α-synuclein line at the 9 and 10-day time points at P < 0.05