RT-PCR of total RNA from normal and HLTF-overexpressing K562 cells.

RT-PCR conditions are described in the “Materials and methods” section. After 18 (left) or 22 (right) cycles, the PCR products were run on a 6% polyacrylamide gel for 2.5 hours, dried, and exposed to x-ray film. The data are representative of 3 independent experiments. K562 + HLTF transfection-1 and transfection-2 are the 2 pools of K562 cells stably transfected with pCIneo containing HLTF in 2 independent transfections. The exposure times were adjusted for each set of PCR to obtain bands of optimum intensity. The autoradiogram exposure times are as follows: 16 hours at room temperature (RT) for β-globin, 12 hours at −80°C for EKLF, 5 hours at RT for ε-globin, 15 minutes at RT for γ-globin, 1 hour at RT for β-actin, and 0.5 hours at RT for GAPDH.