Extended Data Fig. 7: Regulation of AMPARs and GABA_AR synaptic levels by CMA.

a–f, Representative images of MAP2⁺GluA1⁺ neuronal processes (a) and GluA1⁺ neuronal soma (c) in L2AKO and control (Ctrl) primary neurons (obtained from female and male pooled brains) following incubation with CTZ (25 μM, 1 h prior to collection) or AMPA (100 μM, 30 min prior to collection) and Leupeptin (200 μM, 24 h prior to collection). Inset: higher magnification of boxed areas. Scale bars: 50 μm (lower magnification panels) or 20 μm (insets). Quantification of GluA1 lysosomal degradation in processes (b) or soma (d,e) upon addition or not of CTZ and changes in GluA1 levels in soma following CTZ treatment (f). n = 28–48 fields from 4–5 independent experiments per group. g–i, Representative images of MAP2⁺Bassoon1⁺GABA_AR β1⁺ (g) and MAP2⁺vGluT1⁺GABA_AR β2,3⁺ (h) neuronal processes and quantification (i,j) of total levels (left and middle panels) and lysosomal degradation (right panels) in L2AKO and Ctrl primary neurons (from female and male pooled brains) following incubation with Leupeptin. Insets: higher magnification. Scale bars: 50 μm (lower magnification panels) or 5 μm (insets). The dotted lines in b, e, f, i(right) and j(right) indicate no changes relative to untreated samples (NL or CTZ/untreated = 1). From left to right in each graph: n = [30, 40] (b left), [39, 40] (b right), [31, 32, 31, 37, 38, 37] (d), [30, 34] (e), [32, 38] (f), [16 each] (i), [16, 20] (j left, j middle), [15, 14] (j right) fields from 4–8 independent experiments per group. Two-way ANOVA with Tukey’s multiple comparison test (d) or two-tailed unpaired Student’s t-test (b,e,f,i,j) are used. Data are presented as mean values ±SEM.

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