Research Paper Volume 2, Issue 12 pp 924—935

DNA damaging agents and p53 do not cause senescence in quiescent cells, while consecutive re-activation of mTOR is associated with conversion to senescence

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Figure 11. Serum stimulation is required for senescent morphology in nutlin-treated MEL-10 cells. A. MEL-10 cells treated with 2.5 μM nutlin-3a in the presence (Nutlin) or absence (No serum+Nutlin) of serum for 3 days were stained for beta-Gal and microphotographed (left panels). In parallel, 10% serum was added to a replicate well. After 3 days of serum stimulation cells were stained for beta-Gal and microphotographed (right lower panel). Bars - 50 micron.

B. MEL-10 cells treated with 2.5 μM Nutlin-3a (N) in the absence or presence of 10% serum for 24 hr were lysed and subjected to immunoblotting, as indicated. Treatment with 10 nM rapamycin (R) is used as a control for mTOR inhibition.