FIGURE 5.

Akt signaling in SODD^−/− MEFs is reduced. A, primary SODD^+/+ or SODD^−/− MEFs were serum-starved and either left untreated or stimulated with EGF (40 ng/ml) for the indicated times. Total cell lysates were immunoblotted with antibodies specific for either phospho-Akt-Ser⁴⁷³ (top), phospho-Akt-Thr³⁰⁸ (middle), or Akt (bottom) antibodies. Shown are representative immunoblots from two independent experiments. B and D, primary SODD^+/+ or SODD^−/− MEFs were treated as in A, and the level of phospho-Akt-Ser⁴⁷³ (B) or phospho-Akt-Thr³⁰⁸ (D) was determined by densitometry, standardized to Akt protein control for each time point. C and E, primary SODD^+/+ or SODD^−/− MEFs were treated as in A, and the level of phospho-Akt-Ser⁴⁷³ (C) or phospho-Akt-Thr³⁰⁸ (E) at 5 min was determined by densitometry, standardized to Akt loading control, and represented as a relative value to that detected in SODD^+/+ lysates (arbitrarily set as 1). Bars, mean ± S.E. (error bars) of two independent experiments. F and G, primary SODD^+/+ or SODD^−/− MEFs were treated as in A, and the level of phospho-Foxo1-Ser²⁵⁶ was determined by densitometry standardized to Foxo1 protein control for each time point. Shown are representative immunoblots from two independent experiments.