Figure 2.

Effect of insulin or H₂O₂ on the PDK-1 recruitment to the membrane fraction (A) and Akt activation (B and C) in SK-N-SH and SK-N-BE(2) cells. Membrane fractions (A) or whole cell lysates (B and C) (50 μg of protein) prepared from cells treated with insulin (100 nM), H₂O₂ (1 mM), or insulin plus H₂O₂ for 10 min were analyzed with anti-PDK-1 (A), or anti-phospho-Akt (Ser473) (B and C). The same blots were reprobed with anti-actin (A) and anti-Akt (B and C), respectively, to confirm equal protein loading.