Research Paper Volume 1, Issue 1 pp 58—67

Dual regulation of TERT activity through transcription and splicing by ΔNP63α

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Figure 5. ΔNp63α overexpression modulated the overall telomerase activity and induced a S-β-gal activity. The mouse keratinocytes from the p63-/+ mice (samples 1, 3, 5, 7, 9) and ΔNp63αtransgenic mice (samples 2, 4, 6, 8, 10) were treated with the control media (samples 1 and 2) or Sirtinol (100 μg/ml for 24h, samples 3 and 4) or transfected for 72h with shRNA against SIRT1 (samples 5 and 6), p53 (samples 7 and 8) and Sp1 (samples 9 and 10). (A) Telomerase activity. Telomerase activity was determined by the TRAP assay using 1 μg of protein extract obtained from 2x105 cells. Quantitative analysis was done using Molecular Dynamics densitometer and ImageQuant software. The intensity of the positive control lane was taken as 100%. The experiment was repeated three times, and error bars represent mean ± S.D. (B) S-β-gal activity. The S-β-gal activitywas measured using a senescence kit.