Figure 1

General scheme for synchronization and treatment of TM6 cells with Se-methylselenocysteine (MSC) including the collection times. The TM6 cells were plated at a density of 6.6 × 10³ cells/cm² in either 100 mm dishes or six-well plates. After 48 hours of growth the cells were starved in DMEM/F12 medium without growth factors and serum (minimal medium) for a further 48 hours. The cells were released from starvation with DMEM/F12 medium containing growth factors (5 ng/ml epidermal growth factor (EGF) and 10 μg/ml insulin) and serum (2% adult bovine serum). After a further 6 hours MSC was added at a final concentration of 50 to 400 μM (depending upon the experiment) to one set of cells. Untreated cells served as controls. The cells were collected after starvation (0 hours), then at 6 (before the addition of MSC), 9, 12, 16 and 24 hours time-points.