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. 2001 Jul 3;98(14):7982–7987. doi: 10.1073/pnas.131211098

Figure 1.

Figure 1

Expression of mutant-template telomerase RNA. (A) The 11-nt template region is located near the 5′ end of the human telomerase RNA. Ectopic expression of the hTER construct leads to the production of three detectable species (green bars), in addition to the endogenous hTER (blue bar). The template sequences of WT, AU5, U11, and 49A telomerase RNAs are indicated below. (B) Northern blotting analysis of various WT-hTER- and MT-hTer-expressing clonal lines. The 451-nt endogenous hTER is present in every cell line examined (blue bar). Parental lines express only this endogenous species. Arrow, the mature functional MT- hTer or WT-hTER transcript. Broad band at ≈1 kb, polyadenylated species transcribed from the introduced hTER/hTer construct. Lower band at ≈300 nucleotides, degradation product from the introduced hTER/hTer construct. LNCaP clonal lines: lanes 1–4, WT-hTER expressing lines R10, R11, R12, and R19; lanes 5–8, U11-hTer expressing lines 11.1, 11.3, 11.6, and 11.19; lanes 9–12, AU5-hTer expressing lines 5.4, 5.5, 5.14, and 5.15. MCF-7 clonal lines: lanes 13–16, WT-hTER expressing lines K2, K3, P1, and A4; lanes 17–20, 49A-hTer expressing lines D2, G4, K6, and P1. (C) TERT is limiting for telomerase RNA levels. Northern blotting analysis of LNCaP clonal lines R10 (WT, lanes 1–3), 11.6 (U11, lanes 4–5), and 5.4 (AU5, lanes 6–8) after transfection with hTERT. Lane 1: clonal line R10-c (vector control); lane 2: clonal line R10–6 (low level of hTERT mRNA expression); lane 3: clonal line R10–3 (high level of hTERT mRNA expression); lane 4: clonal line 11.6–2 (hTERT expression not detected); lane 5: clonal line 11.6–1 (high level of hTERT mRNA); lane 6: clonal line 5.4-c (vector control); lane 7: 5.4–19 (hTERT expression not detected); lane 8: clonal line 5.4–5 (high level of hTERT mRNA).