FIG. 6.

Wortmannin does not inhibit small-t-antigen-induced Akt or PKC λ phosphorylation. 3T3-L1 adipocytes infected with either control virus or small-t-antigen-encoding adenovirus (ST) were starved, pretreated with 100 nM wortmannin for 30 min, and then stimulated with insulin (100 ng/ml) for 5 min (for Akt) or 30 min (for PKC λ). Whole-cell lysates were prepared and analyzed by Western blotting with phospho-specific Akt (serine 473) antibody (top panel), phospho-specific Akt (threonine 308) antibody (second panel from top), or phospho-specific PKC ζ/λ antibody (fourth panel from top). The same lysates were subjected to immunoblotting (IB) with anti-Akt antibody (third panel from top) or anti-PKC ζ/λ antibody (bottom panel).