Fig. 4. Time-dependent changes of ATG proteins expression following ischemia/reperfusion injury.

Immunoblotting of A ATG12-ATG5 conjugate and B ATG4 showing a significant decrease of the indicated proteins in the ischemic retina after 24 h of reperfusion. C Reduced levels of BECN1 were reported in the retinas subjected to ischemia plus 6 h of reperfusion and associated with the accumulation of a 50 kDa proteolytic fragment (figure box). Note that fragment was detectable only by longer exposure time leading to the saturation of the full-length band (long exp, longer exposure). For each animal, retina from contralateral eye was used as control. 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. (3–4 independent experiments for each group). ^#P < 0.05, ^##P < 0.01 vs. control non-ischemic retina (Student’s t test). C, control non-ischemic retina; I, ischemic eye; MW, molecular weight; Rep time, reperfusion time