Extended Data Fig. 4: The decrease in TCA cycle intermediates is secondary to altered glycolysis in the ATG5 cKO cerebellum.
From: Autophagy regulator ATG5 preserves cerebellar function by safeguarding its glycolytic activity
a,b, Relative profile of TCA cycle intermediates in 1 M (a) and 3 M (b) WT and ATG5 cKO cerebellum. N = 5 per genotype. Multiple unpaired two-sided t-Tests with linear Benjamini, Krieger and Yekutieli correction. c,d, ECAR rates during Seahorse XF glycolysis stress test in WT and ATG5 cKO primary cerebellar cells cultured either in 10 or 30 mM glucose. WT10 mM: 29 wells from N = 6, ATG5 cKO10 mM 22 wells from N = 5, WT30 mM: 29 wells from N = 6, ATG5 cKO30 mM 17 wells from N = 4 with ± 20 000 cells per well. Two-way ANOVA followed by Holm-Šidák multiple comparisons (WT10 mM vs WT30 mM: p < 0.0001; ATG5 cKO10 mM vs WT10mM: p = 0.0005, WT30 mM vs ATG5 cKO30 mM: p = 0.024, ATG5 cKO30 mM vs ATG5 cKO10mM: p < 0.0001). e, Enrichment of 13C-glucose in the glucose-6-P M + 6 fraction in acute cerebellar WT and ATG5 cKO slices at 3 M (N = 10 for WT, N = 8 for ATG5 cKO mice). cKO values are normalized to WT. One-tailed unpaired t-Test. f, Analysis of the metabolic flux of 13C-glucose into TCA cycle intermediates in acute cerebellar WT and ATG5 cKO slices at 3 M. N = 10 mice for WT, N = 8 for ATG5 cKO. cKO values are normalized to WT. One-tailed unpaired t-Test. 1 M and 3 M indicate 1 and 3 months of age, respectively. 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.
