Fig. 2.

Consequences of GSE24.2 expression on telomerase activity of F9 and FA353V cells. (a) F9 and F9A353V cells were transfected with pLNCX, pDKC and pGSE24.2 (10 µg DNA per 10⁶ cells). Telomerase activity was determined with the TRAP assay kit. Different extract dilutions are presented for each TRAP assay as indicated by the triangles. Asterisks indicate the samples that showed important differences in relation to the pLNCX transfected cells. The experiments were repeated 3 times, with similar results. (b) Levels of telomerase complex components in F9 and F9A353V cells: F9 and F9A353V were transfected with either pLNCX, pDKC or pGSE24.2. mTERT mRNA and mTR levels were determined by RT-PCR. β-actin mRNA was used as control. The experiments were repeated 3 times, with similar results. (c) F9 and F9A353V cells were co-transfected with the hTERT-luc reporter vector (1 µg per million cells) and pLNCX, pDKC and pGSE24.2 expression vectors. After 24h, the cells were processed and the luciferase activity determined. The activity obtained is represented as fold induction over the pLNCX control transfection (d) F9 and F9A353V cells were co-transfected with the pXP3.2myc promoter (1 µg per 10⁶ cells) and pLNCX, DKC and GSE24.2. After 24h, the cells were processed and the luciferase activity determined. (e) F9 and F9A353V cells were co-transfected with the hTR-luc reporter vector (1 µg per 10⁶ cells) and pLNCX, pDKC and pGSE24.2 expression vectors. After 24h, the cells were processed and the luciferase activity determined as described in panel d. The experiments were repeated 3 times, with similar results.