Extended Data Fig. 3: Consequences of CMA blockage on the synaptic proteome.
a, Comparative proteomics of whole-brain ex vivo slices from 9 months old male and female control (Ctrl) and L2AKO mice incubated with or without lysosomal protease inhibitors (20 mM ammonium chloride and 400 μM leupeptin, NL) for 20 h. Timeline for ex vivo slice culture and NL incubation of the brain slices is shown. b,c, Representative immunoblot (b) and quantification (c) of lysosomal degradation of LC3-II (changes in protein levels upon NL addition relative to (left) or minus untreated (right)). n = [6] mice per group (c). Ponceau staining is shown as loading control. The dotted line in c indicates no changes relative to untreated samples (NL/untreated = 1). d, Proteins detected by mass spectrometry as increased (>1.5-fold, orange) or reduced (<0.8- fold, blue) in L2AKO females or male brains relative to controls. e,f, GO terms (e) and STRING plots (f) of the pathways enriched in proteins with increased steady-state levels in L2AKO brains compared to Ctrl female and male brains. All GO terms are statistically enriched with P < 0.05. Data are presented as mean values ±SEM. False discovery rate (FDR) test was used to calculate values in e.
