Figure 5.

Insulin-stimulated serine phosphorylation of Akt in isolated microvessels ex vivo. Isolated microvessels were stimulated without or with insulin (2 and 100 nM) for 20 minutes at 37°C (n = 3; each sample of microvessels was derived from 2 rats). Tissues were homogenized in lysis buffer as described in Methods. Lysates (100 μg protein) were separated with 8% SDS-PAGE and transferred to nitrocellulose membrane. The membranes were blotted with anti-phospho-(serine 473)-Akt antibody, viewed using an ECL kit, and quantified with a densitometer. *P < 0.05, **P < 0.01, lean vs. obese treated with insulin at the same concentrations (n = 3; each sample of microvessels was derived from 2 rats).