Figure 3.

Macrophages from p47^phox-/- mice have decreased Akt1 and p38 MAPK activation, but not Erk activation. BMM were derived from p47^phox-/- or wild-type littermate mice (WT) by growth in the presence of M-CSF (20 ng/ml) for 5 d. The cells were serum-starved overnight, then stimulated with M-CSF (10, 50, or 100 ng/ml) for 5 min or left unstimulated. (A) Akt1 activation was assessed using equal amount of protein by Western blotting with antibodies to pAkt^Thr308 (upper panel) and pAkt^Ser473 (middle panel). Membranes were reblotted for total Akt1 (lower panel). (B) Western blot analysis using phospho-p38 MAPK antibody and equal loading was confirmed by p38 MAPK antibody. (C) Erk phosphorylation was detected using pErk 42/44 antibody. Equal loading is shown using Erk2 antibody. Shown are representative data from macrophages obtained from six independent mice.