Extended Data Fig. 3: Increased autophagy and lipid metabolism in metabolically stressed optic nerves.
From: Oligodendroglial fatty acid metabolism as a central nervous system energy reserve
a, Left: silver stained gels of optic nerve lysates from 2 month old wildtype mice (both genders), prepared after 24 h incubation in 10 mM glucose or 1 mM glucose (two nerves pooled per lane; one lane is equivalent to one sample). N = n = 5 for both conditions. Right: optic nerve lysates prepared after 16 h in 10 mM glucose or 0 mM glucose. Note the lack of major protein degradation. N = n = 5 for both conditions. b, Relative abundance of selected proteins in optic nerve lysates after 24 h in 1 mM glucose (left, N = n = 5) or 16 h in 0 mM glucose (right, N = n = 5). Note that enzymes of glucose and lipid metabolism show only moderate changes in abundance. Autophagy related proteins are increased in the presence of 1 mM glucose only, indicating a requirement of glucose for RNA synthesis and protein expression (N = n = 5, two technical replicates each; moderated t-statistics (more details in methods section)); Statistical significance (q-value) depicted on the right side of each panel. c, d, Western blots of lysates from wildtype optic nerves, incubated in 10 mM or 0 mM glucose for 16 h (age 8-12 weeks old, N = n = 5 for each condition) (c) and quantification of ACAT1 and BDH1 (d). Normalized to protein input (mean ± SEM, unpaired two-tailed t-test). e, Cell survival of 24 h glucose-deprived optic nerves from TFEB cKO mice (N = n = 4) and controls (N = n = 4; age 8-12 weeks). Images from longitudinal sections were stained with PI and DAPI. f, Quantified data from (e). There is no difference of cell survival (mean ± SEM, unpaired two-tailed Welch’s t-test). N and n indicate the total number of independent samples for each condition and the total number of independent experiments, respectively.
