Fig. 2: Non-mitochondrial autophagic flux is preserved in Drosophila expressing human wild-type or mutant α-synuclein.
a Confocal images of Keima emission in indirect flight muscle of 4-week-old control flies (CTRL1: w1118;;UAS-Keima,mef-2-GAL4/+) at 458 nm (green) and 543 nm (red) excitation. Arrowheads indicate ‘acidic’ puncta with high 543/458 ratio values. b Confocal images of Keima-expressing indirect flight muscle of 4-week-old CTRL1 labeled with LysoTracker (100 nM), showing colocalization of ‘acidic’ Keima puncta with lysosomes (arrowheads). Arrows indicate examples of lysosomes without Keima signal. Additional images of colocalization of high 543/458 ratio Keima puncta with LysoTracker are shown in Supplementary Fig. 1. c High Keima (543/458) ratio signal of 1- and 4-week-old CTRL1 or Atg1K38A-overexpressing indirect flight muscle tissue. d Quantification of (c). High Keima (543/458) ratio area/ total cell area was quantified as an index of non-mitochondrial autophagic flux (n = 5−6 flies per condition, from at least 3 different crosses). One-way ANOVA with post hoc Tukey’s test. *P < 0.0001 compared with 1-week-old Atg1K38A flies and 4-week-old CTRL1 flies. #P < 0.0001 compared with 4-week-old Atg1K38A flies. e High Keima (543/458) ratio signal in indirect flight muscle of 4-week-old CTRL1 and CTRL2 (w1118;;UAS-Keima,mef-2-GAL4/UAS-smGdP) flies and flies expressing wild-type (w1118;;UAS-Keima,mef-2-GAL4/UAS-SNCA WT) or mutant A53T α-synuclein (w1118;;UAS-Keima,mef-2-GAL4/UAS-SNCA A53T). f Quantification of (e) (n = 5 flies per condition, from at least 3 different crosses; P = 0.16, one-way ANOVA with post hoc Tukey’s test). Error bars represent SEM. Scale bars, 10 µm.
