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. 2001 Jan 16;98(2):646–651. doi: 10.1073/pnas.98.2.646

Figure 2.

Figure 2

Telomerase activity. (A) 20- and 200-ng aliquots of cellular extracts were assayed for the presence of telomerase activity by the telomeric repeat amplification protocol (TRAP) assay. Extracts (200 ng each) were heat-treated (HI) to inactivate telomerase and assayed as a negative control. An internal PCR standard was included in each PCR to demonstrate that an absence of telomerase did not result from PCR inhibitors in the cell extracts. (B) Telomerase activity measured by the TRAP assay was detected in cellular extracts (20 and 200 ng) from HMF3 and HMME7 cells containing ectopic hTERT and U19tsA58 LT cultured at both 33.5°C (permissive temperature for tsA58 LT) and at 39.5°C (nonpermissive temperature) for 7 days. Activity was not detected in HMF3 and HMME7 cells containing U19tsA58 LT alone. (C) Southern blot showing TRF lengths for the fibroblast cultures HMF3 at p3 (lane 1), p12 (lane 2), p18 (lane 3), p24 after early transduction of tsLT (lane 4), p23 after late transduction of tsLT (lane 5), and p17 after both early (lane 6) and late (lane 7) transduction of hTERT. Numerical values for mean TRF lengths of these and other cultures are shown in Table 1.