Effect of TUCAN down-regulation on cisplatin sensitivity in H460 cells. A) Real time RT- PCR experiment and B) Western blot analysis showing the silencing of TUCAN expression in H460 cells stably transfected with pSUPERretro TUCAN3 (pSrpTUCAN3). As control, a construct containing the reverse orientation of the RNA interference sequence, pSUPERretroTUCAN-control (pSrp control), was used. The mean ± SD of three independent experiments is shown, * – non-specific band. C) Cisplatin- and etoposide-induced apoptosis in H460 cells and derived transfectants. The percentage of cells in the sub-G1 fraction was taken as the apoptotic population. The results are depicted as the means of three experiments; bars, SD.D) activation of caspase-9 was measured by the cleavage of the fluorogenic substrate LEHD-AFC in H460 cells and derived transfectants exposed to cisplatin.