FIG. 6.
C2-ceramide does not inhibit tyrosine phosphorylation of IRS-1. 3T3-L1 adipocytes were treated with C2-ceramide (C2) (100 μM) or C2-dihydroceramide (C2H2) (100 μM) for 2 h prior to solubilization, with insulin (20 nM) present for the last 10 min. IRS-1 was immunoprecipitated (IP), separated on SDS-polyacrylamide gel electrophoresis gels, and transferred to nitrocellulose. Western blots (WB), were probed with antiphosphotyrosine antibodies (αpTyr), and the blots shown are representative of two independent experiments.