HeLa cells were grown in DMEM medium supplemented with 10% fetal bovine serum at 37°C and 5% CO2. siRNA-mediated knockdown of either EGFP(control), and hMTR4 (SKIV2L2) were performed using 22 nM of siRNA and Lipofectamin2000 (Invitrogen) as transfecting agent. A second hit of 22 nM siRNA was given after 48 h. Cells were harvested an additional 48 h after the second hit, and protein depletion was verified by western blotting analysis as described elsewhere48. The following siRNA sequences were used:
