Induction of S and T phosphorylation of Akt by G-CSF.
Parental Ba/F3 cells or Ba/F3 cells transfected with the WT G-CSFR or the Δ716 G-CSFR were untreated (0) or stimulated with 100 ng/mL G-CSF (+) for 10 minutes, then lysed and immunoprecipitated with anti-Akt antibody. (A) Immunoblotting with anti–phospho-Akt (S473). (B) The blot in (A) was stripped and reblotted with antibody recognizing Akt phosphorylated at T308. (C) The blot in (B) was stripped and reblotted with anti-Akt to assess for equivalent protein loading. Arrows indicate the phosphorylated forms of Akt. Peroxyvanadate (pVO4)-stimulated Δ716 cells are shown as a positive control. Δ716 cells immunoprecipitated with normal sheep serum (IgG) instead of anti-Akt antibody represent a negative control.
