Figure 6. Effect of isoform-specific inhibitors on insulin signalling in J774.2 cells.

Overnight-starved J774.2 cells were incubated for 5 min with wortmannin (wrt; 100 nM), PIK-75 (50 nM), TGX-221 (50 nM), IC87114 (100 nM) or DMSO and stimulated or not with insulin (100 nM, 10 min). Whole-cell lysates were then analysed by Western blotting using antibodies recognizing phospho-Ser⁴⁷³ (pS473) of PKB. A representative Western blot is shown in the upper panel and quantification is presented below. The relative stimulation of insulin over the basal level was taken as 100% and all values are calculated as a percentage of this. Results are means±S.E.M. for three independent experiments each performed in duplicate. ***P<0.001.