FIG. 10.

Regulation of AMPK phosphorylation in U251MG cells. U251MG cells were either serum starved for 3 days and released in 10% serum or treated with 10 μM PD98059, 20 μM LY294002, 10 μM carbonyl cyanide m-chlorophenylhydrazone (CCCP), and 2 mM 5-aminoimidazole-4-carbozamide-1-β-4 ribofuranoside (AICAR) in the presence of 10% serum overnight. Cell lysates were subsequently collected to perform immunoblotting using antibodies against AMPK and phospho-AMPK (T172), followed by horseradish peroxidase-conjugated secondary antibodies, and visualized by the ECL reaction.