Figure 2

chTERT in-frame isoforms are non functional in the in vitro telomerase assay. (A) Schematic diagram of the 3 reconstituted in-frame isoforms of chTERT inserted into pcDNA under the control of the T7 promoter. Amino acids deleted or inserted in each isoform are indicated and those belonging to the RT motif are underlined. Plasmids were used to produce recombinant in vitro-translated proteins that were incubated in the presence of in vitro-transcribed vTR or chTERT before the TRAP assay. (B) The histogram shows telomerase activity relative to the value obtained with vTR in the presence of the full-length constitutively spliced chTERT transcript arbitrary set at 100%. (C) Histograms show telomerase activities for a mixture (1:1) of in vitro-translated full-length chTERT and the indicated isoform in the presence of vTR or chTR. The values obtained with the mixture of full-length chTERT and isoform are expressed relative to the value for the full-length chTERT, arbitrarily set at 100%. The histogram shows the mean and standard deviation obtained from 3 biological analyses.