In vitro model of ischemia. (A) Analysis of cellular ATP during OGD. Reduction in ATP becomes significant after 2 h. In the presence of glucose, ATP levels remain unchanged. OD, oxygen deprivation. (B) Decreased intensities of TMRM and J-aggregated form of JC-1 (red) probes under OGD indicate ΔΨm depolarization. (C) Dissimilar to OD, only a minor transient elevation in Epas1 (also known as HIF-2α) levels is seen under OGD (arbitrary units (a.u.)). (D) Characteristic OGD-induced changes in phosphorylation of the proteins involved in regulation of translation. A minor increase in phosphorylated AMPKα under OD suggests a smaller effect of this condition on AMPK signaling. Scale bar is 50 μm. Asterisks demonstrate significant difference. Error bars show standard deviation (A,C,D) or a range in TMRM and JC-1 intensities (B), calculated for 40 to 50 randomly selected cells.