Extended Data Fig. 8: GABAergic ATG5 deletion does not induce apoptosis or alter MG-modified protein levels in cerebellar interneurons.

a-c, Fluorescence-based analysis of MG-modified proteins in PC dendrites (b) and molecular cell layer interneurons (c) in WT and ATG5 cKO mice. Scale bar, 50 µm (Inserts, 10 µm). N = 4 for 1 M and N = 5 for 3 M animals. Two-way ANOVA followed by Holm-Šidák multiple comparisons. In b WT^3M vs cKO^3M: p = 0.027. d,e, Fluorescence-based analysis of D-Serine in molecular cell layer interneurons from WT and ATG5 cKO mice. Scale bar, 25 µm. N = 4 mice for 1 M and N = 5 for 3 M. Two-way ANOVA followed by Holm-Šidák multiple comparisons. f,g, Representative confocal images of WT OTCs cultured in media containing either vehicle, LPA or LPA+Fer1. Scale bar, 100 µm. See quantification in Fig. 6n. 15 images for control, 17 images for LPA and 17 images for LPA + Fer1, from N = 3 independent experiments. g, Representative confocal images of WT OTCs cultured in media containing either vehicle, D-Serine or D-Serine+Fer1. Scale bar, 100 µm. See quantification in Fig. 6o. 14 images for control, 22 images for D-Serine, 17 images for D-Serine + Fer1, from N = 4 independent experiments. h,i, Fluorescence-based analysis of cleaved Caspase-3 levels in PCs from WT and ATG5 cKO mice. Scale bar, 200 µm. N = 3 per genotype for 1 M and N = 4 per genotype for 3 M. Two-way ANOVA followed by Holm-Šidák multiple comparisons. j,k, GCAMP7f-based analysis of untreated WT PCs, LPA-treated WT PCs and ATG5 cKO PCs. (159 PCs for WT, 79 PCs for WT^LPA and 117 PCs for ATG5 cKO, from N = 3 per genotype). One-way ANOVA followed by Holm-Šidák multiple comparisons. 1 M and 3 M indicate 1 and 3 months, respectively. m.g.v.-mean gray value. Squares in Fig. S8a indicate regions magnified. All graphs show mean ± SEM. n.s.—non-significant; * indicates P ≤ 0.05; ** indicates P ≤ 0.01; *** indicates P ≤ 0.001; **** indicates P ≤ 0.0001.

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