Extended Data Fig. 5: pAMPK levels and GLUT1/3/4 levels in the ATG5 cKO cerebellum.
From: Autophagy regulator ATG5 preserves cerebellar function by safeguarding its glycolytic activity
a, 2-NBDG uptake in PCs from WT OTCs from 1 independent experiment. b, 2-NBDG uptake in molecular cell layer interneurons in WT (290 cells from N = 6) and ATG5 cKO (242 cells from N = 4) OTCs. Two-tailed unpaired t-Test. c, Ratiometric analysis of Laconic and AUC in molecular cell layer interneurons in WT (158 cells from N = 5) and ATG5 cKO (134 cells from N = 4) OTCs at DIV21. Two-tailed unpaired t-Test. d,e, Protein levels of AMPK and its phosphorylated form (pAMPKα-Thr172) in cerebellar WT and ATG5 cKO lysates at 3 M. N = 4 per genotype. Two-tailed unpaired t-Test. f-k, Representative confocal images and analysis of WT and ATG5 cKO cerebellum, immunostained for GLUT1 (N = 3 for 1 M per genotype and 3 M WT and N = 5 for 3 M ATG5 cKO animals), GLUT3 (N = 4 per genotype for 1 M and N = 3 for 3 M) and GLUT4 (N = 3 for 1 M and N = 4 for 3 M animals per genotype). Scale bar, 50 µm. Two-way ANOVA followed by Holm-Šidák multiple comparisons. l,m, Fluorescence analysis of hexokinase II (HK2) in PC soma of WT and ATG5 cKO cerebellum at 1 M. Scale bar, 50 µm (Inserts, 10 µm). N = 4 mice per genotype. Two-tailed unpaired t-Test: p = 0.017. n, Slc2a2 mRNA levels in WT and ATG5 cKO cerebellum at 3 M. mRNA levels of each genotype were normalized to Gapdh. N = 4 mice per genotype. One-tailed unpaired t-Test. o, Ponceau S-stained membrane used for Western Blot shown in Fig. 4n (representative image from N = 4 independent experiments). 1 M and 3 M indicate 1 and 3 months, respectively. m.g.v.-mean gray value. 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.
