Figure 1

Dephosphorylation/inactivation of Akt and Bad and PI-3K in Caco2. Caco2 cells were exposed to 10⁻⁷ mol/L testosterone-HSA for the indicated time periods. The ratio of the cellular content of the phosphorylated residues (Thr 308) versus the total isoform of Akt (A, B) and the phosphorylated residues (Ser 136) versus the total isoform of Bad (C) were measured in cell lysates by Western blotting using specific antibodies for each form and were normalized to the corresponding control. Blots show a representative experiment, the numbers below each lane correspond to the mean values ± SE from three independent experiments (*P < 0.05; **P < 0.01) indicating the fold-increase in the phosphorylation level for the indicated time point normalized to the controls. (B) Control experiment showing variation of p-Akt levels over time (representative blot from two distinct experiments). (D) Following cell lysis equal amounts of total lysates were immunoblotted (IB) with a specific antibody against phospho-PI-3K (p85) and total PI-3K. Immunoblots were analyzed by densitometry. The intensity of phospho-PI-3K (p85) was normalized to the intensity of the corresponding total PI-3K band. Blots show a representative experiment, whereas the relative fold-increases are indicated as mean values ± SE from three independent experiments of PI-3K phosphorylation with that of untreated cells taken as one (*P < 0.05; ***P < 0.001).