At the end of the behavioral test, 5U/kg insulin was injected and 20 min later the hypothalamus, hippocampus and amygdala were removed. Tissue lysates, 20 μg per lane, were subjected to Western blot analysis with antibodies specific for PS473-AKT or total ERK. There were 10 mice in IES with escape failure group (IES+EF group) and 10 mice in sham group. Representative Western blots are presented in A, B and C. White gap in A, B and C indicates grouping of lanes from different parts of the same gel.