Effects of leptin and insulin on phosphorylation of STAT3, MAPK, PKB and GSK3 Rat ARC wedges were incubated for 0, 1, 5, 15 or 30 minutes with 10 nM leptin (A) or 0.1 – 1 nM insulin (B) before cells were lysed and equal amounts of lysate were subjected to SDS-PAGE and transferred to nitrocellulose membrane. The phosphorylated levels of p42/p44 MAPK, PKB, STAT3 and GSK3α/β were detected by immunoblotting with appropriate specific antibodies. The total amount of PKB is also shown. Bands were quantified using densitometry. The values are expressed as relative to the corresponding aCSF control group, and normalized for protein loading. Values represent the mean ± SEM for between 4–6 animals for each time point. * P < 0.05 and ** P < 0.01.
Mirshamsi et al. BMC Neuroscience 2004 5:54 doi:10.1186/1471-2202-5-54