Fig. 1

Generation of the wlz56 dat- 1::lgg-1::mCherry C. elegans line. a. Structure of dat-1::lgg-1::mCherry reporter construct. b. Expression of dat-1::lgg-1::mCherry construct in dopaminergic neurons. The left panel shows the expression pattern of dat-1::GFP, the middle panel shows the expression pattern of dat-1::lgg-1::mCherry, and the right panel shows the merged images demonstrating the overlapping expression. c. Immunoblot showing expression of lgg-1::mCherry, which shows a monomeric band at approximately 37 kD (arrow). d. Levels of the dat- 1::lgg-1::mCherry reporter reflect autophagic flux. Exposure of wlz56 to bafilomycin (100 μg/ml, 72 h) increases fluorescence. Arrows point to puncta of dat- 1::lgg-1::mCherry fluorescence. e. Quantification of the lgg-1::mCherry reporter fluorescence showing increased expression in nematodes treated with 100 μg/ml bafilomycin for 72 h. f. Deletion of Atg5 increases lgg-1::mCherry protein. Immunoblot showing total dat-1::lgg-1::mCherry protein from 50 crossed heterozygous F1 males. Control contains males (F1) from N2 crossed with wlz56 (dat- 1::lgg-1::mCherry) while Atg5 contains males (F1) from Atg5 mutant strain crossed with wlz56. A monomeric 37KD band represents the dat-1::lgg-1::mCherry fusion protein. g Total lgg-1::mCherry band intensities were scanned and quantified lines containing WT or mutated ATG5 (n = 8). Scale bar = 20 μm