Figure 3

Effects of shRNA on telomere length and telomerase activity in Colo 320 cells. A. Representative the concentration-course analysis of telomerase activity, each groups cells were mixed with 1 ml TBA solution for preparation of protein extract and 1 μg protein was subjected to TRAP assay. After hybridization and ELISA procedure, the absorbance of the samples at 450 nm was measured. The cells were subjected to no treatment (control), pGenesil-vector group, 5, 7.5, 10, 12.5 μM shRNA for 48 h. B. Mean telomere restriction fragment length was detected by RT-PCR with different treatments by Southern analysis as described under Materials and methods. Typical Southern blot results for telomere restriction fragments. Locations of the base pair markers on the DNA ladder are indicated along the left side. Significant difference was observed between the mean telomere lengths of the control and shRNA-transfected groups cells. All data were obtained from three independent experiments. Error bars represent means ± SEM. Significantly different from the corresponding control (**P < 0.01, vs control. ##P < 0.01, vs vector).