Effect of Se-methylselenocysteine (MSC) on Akt. Synchronized TM6 cells were treated with 50 μM MSC as described in Fig. 1. For each time point three 100 mm dishes were treated with MSC and pooled for protein content. Equal amounts of protein lysates were loaded on each lane (50 μg) for each time point. (a) Immunoblots were probed with anti-Akt, anti-phospho-Akt (Ser473) and anti-actin as described in the Materials and methods section. The phosphorylation of Akt occurring at 24 hours in control cells was inhibited in the MSC-treated cells. (b) The levels of phospho-Akt in control and MSC-treated cells were normalized with respective actin contents and plotted against various time points.