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. Author manuscript; available in PMC: 2016 Jul 11.
Published in final edited form as: Nat Genet. 1999 Dec;23(4):405–412. doi: 10.1038/70508

Fig. 6.

Fig. 6

Telomerase dependence. a, Telomerase activity. We infected WI-38 (lanes 1,2) or hTERT (hT)-expressing WI-38 (lanes 3–8) cells with control (Lx, lanes 1–4), TIN2 (lanes 5,6) or TIN2-13 (lanes 7,8) retroviruses, selected for virus-expressing cells and prepared cell lysates. We analysed cell lysate volumes equivalent to equal numbers of cells for telomerase activity by TRAP assay. neg, extracts heated to 85 °C before assay. b, TRF length. We infected WI-38 cells at PD 29 (lane 1) with pBabe control (lanes 2,4,6,7) or hTERT-expressing (lanes 3,5,8,9) virus. After five to six PD, we superinfected the cells with viruses expressing LXSN control (Lx; lanes 2,3), TIN2 (lanes 4,5) or TIN2-13 (lanes 6–9). We isolated DNA at the indicated PD levels, and analysed the DNA for TRF length. c, TIN2 does not inhibit telomerase activity in vitro. We prepared extracts from HT1080 cells (lane 2) and mixed equal aliquots with 20 ng GST (lane 4), or 1 (lane 5), 5 (lane 6) or 20 (lane 7) ng GST–TIN2. We incubated the extracts for 10 min at 4 °C before assaying for telomerase activity. pos, positive extract from the assay kit; neg, extract heated to 85 °C.