Fig. 5. Time-dependent modulation of mTOR signaling pathway upon retinal ischemia.

Retinal ischemia was induced in the right eye and mice were killed after 0, 1, 6, or 24 h of reperfusion. For each animal, contralateral non-ischemic retina was used as control. The phosphorylation level of A mTOR (p-mTOR), C AMPK (pAMPK), and D Akt (p-Akt) was studied in whole retinal lysates by western blotting. mTOR activity was indirectly checked by analyzing the state of phosphorylation of its downstream targets A ULK1 and B 4EBP1. Histograms represent the densitometric analysis of the bands normalized to loading control (actin). Dashed lines indicate the baseline expression of the protein in non-ischemic retinas set to 1. Data are reported as mean ± s.e.m. of 3–7 independent experiments for each group. ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 vs control non-ischemic retina (Student’s t test). C, control non-ischemic retina; I, ischemic retina; MW, molecular weight; Rep time, reperfusion time