FIG. 8.

C₂-ceramide inhibits insulin- and PDGF-stimulated Akt phosphorylation in fibroblasts. Total cell lysates were prepared from 3T3-L1 fibroblasts preincubated with C₂-ceramide (C2) (100 μM) for 2 h followed by a 10-min incubation with insulin (1 μM) or PDGF (50 ng/ml). Proteins were separated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose, and Western blots were then probed with anti-phospho-Akt antibodies (αPAkt).