FIG. 4.

Microinjection and treatments of inhibitors and p110caax expression on Gαq expressed 3T3-L1 adipocytes. (A) After 60 h of Gαq-expressing adenovirus infection (MOI = 10), 3T3-L1 cells were injected with antiphosphotyrosine antibody (PY-20), GST-BARK, or sheep IgG as a control, prior to 3 h of insulin stimulation. Fixed cells were stained with rabbit anti-GLUT4 antibody and incubated with TRITC-conjugated anti-rabbit IgG antibody and AMCA-conjugated anti-sheep IgG antibody, as described in Materials and Methods. (B) After 60 h of Gαq-expressing adenovirus infection (MOI = 10) with or without p110caax-expressing adenovirus coinfection (CAAX), 3T3-L1 adipocytes were incubated with 300 nM wortmannin (Wort) or 0.1% DMSO vehicle for 60 min and with insulin (100 ng/ml) for 50 min or left untreated. 2-[³H]deoxyglucose uptake was measured as described in Materials and Methods. The data are means and standard errors from three independent experiments.