Figure 2

Influence of AICAR on AMPK phosphorylation and apoptosis in CT-2A cells. CT-2A and astrocyte cells were treated with AICAR (0.5 and 1.0 mM) in glucose, glutamine and serum containing media for 24 hours. a) Cell lysates were probed with specific phosphorylated and non-phosphorylated antibodies for western blot as indicated. Dose dependent increase in AMPK phosphorylation was evident in CT-2A and astrocytes but caspase-3 and PARP cleavage was noted only in AICAR treated CT-2A cells. b) Cells were treated with 1.0 mM AICAR in glucose, glutamine and serum containing media for 24 hours and treated with Annexin V and PI stain as described in methods. Labeled cells were analyzed by flow cytometry. 8–10% apoptotic cells were noted in CT-2A cells treated with AICAR compared to untreated cells. No significant cell-death was noticed in treated astrocytes. All experiments were done in triplicate.