Fig 1: Imbalance between p-AMPK/autophagy and mTOR signaling in CrT–/y neonates.(A) EM showed more membrane whorls (arrows) and large lucid cytoplasm (pink-colored) in the neocortical neuropil of 3-day-old CrT–/y mice, particularly after 12 hours of starvation/separation from the dam (n = 3 for each condition). The numbering 1–4 for the indicated condition is used in all panels of this figure. Scale bar: 1 µm. (B) Immunoblot detection of AGAT and GAMT in P3 CrT+/y and CrT–/y mice (n > 3 for each). Note the expression of AGAT and GAMT in P3 mouse brains with or without starvation, but not in the adult brain. (C) LC-MS measurement of the brain ATP, Cr, and PCr levels in P3 CrT+/y and CrT–/y neonates with or without starvation (n = 5 for each group). (D and E) Immunoblotting analysis and quantification of the p-AMPK/autophagy and mTOR signaling pathways in P3 CrT+/y and CrT-/y mouse brains, with or without starvation as indicated (n = 4 for each). The violin plots in E are representative of 3 independent experiments, with n = 4 biological replicates; all other data are shown as mean ± SEM. All statistical analyses were performed using 2-way ANOVA followed by Tukey’s multiple comparison post hoc test.