Figure 1.
E6-AP expression is required for telomerase induction by HPV 16E6. (A) Northern blot. Three shRNA constructs targeting E6-AP (esh1,2,3) were transduced into HFKs and RNA was harvested to examine levels of E6-AP message. 36B4 is a loading control. Relative expression levels are presented normalized to 36B4 and the pB empty vector control. (B) Western blot. HFKs expressing the E6-AP shRNA constructs were subsequently transduced with LXSN empty vector (-) or LXSN-16E6 (+). p53 levels were examined by Western blot. Actin is a loading control. (C) TRAP assay. Extracts from the same cells shown in B were assayed for telomerase activity. HeLa cells are a positive control lysate. CHAPs is a lysis buffer negative control. TSR8 is a synthetic template of eight telomeric repeats used as a PCR positive control. (D) RT-PCR. Expression of hTERT RNA was examined by RT-PCR of RNA extracts. 36B4 is a loading control.