Fig. 1.
Western blot analysis of macrophage lysates from p110γ+/+ and p110γ−/− mice. (a) Lysates of BMDMs exposed (15 min at 37°C) to various agonists after 24 h of serum deprivation were separated by SDS/PAGE and immunoblotted with phosphospecific Akt (Ser473) antibody (Upper). Separate but identical blots were probed with nonphosphospecific Akt antibody to demonstrate equal loading of lanes (Lower). Agonist concentrations were oxidized LDL, 1 and 10 μg/ml; fMLP, 10 μM; IL-8, 10 ng/ml; MCP-1, 20 ng/ml; lysophosphatidic acid, 10 μM; C-reactive protein, 10 μg/ml; C-peptide, 1 nM; IGF-1, 20 nM. (b) Cultured BMDMs were exposed to angiotensin II at the concentrations indicated, for 5 min at 37°C before cell lysis, in the presence (20 nM) or absence of wortmannin. wt, wild type; ko, p110γ−/−.