Figure 6
Effect of Deptor KD on cell cycle regulation. Control and Deptor KD myoblasts were grown in DMEM supplemented with 10% FBS overnight to 50–60% confluency. Myoblasts were then serum starved for 18–24 h in serum-free DMEM to arrest them in the G1 to G0 phase. Serum-starved myocytes were then released from the cell cycle arrest by the addition of fresh media containing 10% FBS. At 16 h, these proliferating cells were fixed. (A) Bar graph shows the percent of cells in the S-phase of the cell cycle in serum-starved cells (0 h) and after addition of serum (16 h); mean ± SE; n = 5–6 for each condition. *P < 0.05, compared with time-matched control values. (B) Western blot of samples (normalized to total protein content for loading) as described above. Blots were probed with total Deptor antibody (top band) and with phosphospecific antibody to detect phosphorylation on S807/S811 of pRb protein (middle band). The lower band was probed with β-tubulin to confirm equal loading.