Research Paper Volume 3, Issue 11 pp 1078—1091

Yeast-like chronological senescence in mammalian cells: phenomenon, mechanism and pharmacological suppression

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Figure 10. Rapamycin prevents CS of a few indicator cells by indirect effects on abundant cells. A mixture of 2000 HCT116 cells (indicator, non-green cells) and 80000 HT-p21-9 (GFP-expressing, green cells) were plated in 96-well plate with 50 μg/ml IPTG with or without 500 nM rapamycin. After 4 days, equal aliquots of attached cells were re-plated in 60 mm dishes in fresh medium with IPTG to prevent proliferation of HT-p21-9 cells and to allow only viable indicator cells to form colonies. (A) After 7 days, colonies were stained. (B) Microphotograph of live cells before staining. A colony of HCT116 cells with a solitary large HT-p21-9 cell (labeled with GFP) is shown.