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. 2014 Jul 14;111(30):E3062–E3071. doi: 10.1073/pnas.1411370111

Fig. 3.

Fig. 3.

Telomerase inhibition activates E2F1- and p53-dependent PUMA transcription in the absence of p21. Indicated cell lines were treated with either a mismatch oligonucleotide or imetelstat. (A) Fold change in PUMA, BAK, and BAX transcript levels measured by quantitative RT-PCR (qRT-PCR) after 6 wk of treatment. (B) Immunoblot analysis was performed for indicated proteins. (C) PUMA expression was measured in imetelstat-treated HCT116 p21KO cells after E2F1 or control (NS) shRNA knockdown. (D) E2F1 knockdown was confirmed by qRT-PCR. (E) Relative cell viability was measured by trypan blue exclusion assay. (F) Cleaved caspase 3 was measured by immunoblot in mismatch oligonucleotide or imetelstat-treated HCT116 p21KO cells after E2F1 or control (NS) shRNA knockdown. Actin expression was monitored as a control. (G) Annexin V–FITC–positive cells were quantified by FACS analysis in mismatch oligonucleotide or imetelstat-treated HCT116 p21KO cells after E2F1 or control (NS) shRNA knockdown. (H) PUMA expression in imetelstat-treated HCT116 p53KO after p21 or control (NS) shRNA knockdown. (I) p21 knockdown was confirmed by qRT-PCR. (J) Cell viability of HCT116 wild-type and p53KO cells after 6 wk of imetelstat treatment monitored by trypan blue exclusion. *P < 0.01; ***P < 0.0001.