Figure 2

Changes in phosphorylation of PKB, GSK3 and MAPK by inhibition of PI3K Rat ARC wedges were pretreated with 10 μM LY294002 or aCSF for 20 minutes and incubated for 1 minute with 10 nM leptin or 1 nM insulin or aCSF. Equal amounts of protein lysate were subjected to SDS-PAGE and transferred to nitrocellulose membrane. The phosphorylated levels of PKB, and GSK3αβ (A) and p42/p44 MAPK (B) were detected by immunoblotting with appropriate specific antibodies. The total amount of PKB is also shown. Bands were quantified using a densitometer. The values are expressed as relative to the corresponding aCSF control group, and normalized for protein loading. Values represent the mean ± SEM for between 4–5 and 3–5 animals for each time point with leptin and insulin respectively. * P < 0.05 and ** P < 0.01.