Figure 5. The anti-apoptotic effect of IGF-1 is abrogated by the inhibitors of the IGF-1 receptor (AG1024), Akt (LY294002), and Erk-1/2
(U0126). Retinal ganglion cells (RGCs) were cultured in the absence or presence of U0126 (U, 5 µM), LY294002 (LY, 20 µM),
and AG1024 (AG, 10 µM) with or without IGF-1, 200 ng/ml, under normoxia or hypoxia for 24 h. Western blot analysis was performed
to detect (A) activation of total Akt, and phospho-Akt and phosphorylation of Bad at serine 136, (B) activation of total Erk-1/2 and phospho-Erk-1/2, and phosphorylation of Bad at serine 112, and (C) the levels of the active capase-3 subunit. β-actin immunoreactivity from the same gel is shown as the internal control.
Histograms show the results of the relative protein level normalized to β-actin. D: Apoptotic cells were detected TUNEL analyses. Representative results of three independent experiments are shown. The results
are expressed as the means±SEM. *p<0.05, **p<0.01 versus the hypoxia group; #p<0.05, ##p<0.01 versus the hypoxia group; &p<0.05, &&p<0.01 versus IGF-1 treated group.