Figure 2

Adenovirus vectors activate Akt kinase and make cells unresponsive to insulin. Cultured SHSY-5Y cells were treated with 100 nM insulin alone for 5 minutes (Control + Ins), or infected with recombinant adenovirus alone at MOI20 (AdGFP), respectively, and compared to cells similarly treated with insulin but pre-infected with adenovirus 48 h previously (AdGFP + Ins). Cells were harvested for transduction efficiency and total cellular protein, and Western blot analyses were performed with antibodies specific for Akt phosphorylated on serine 473 and total Akt, respectively (lower panel). For quantification, densitometry of the Western blot data were performed (method S.D. < 10%, n = 3).