RLIP76 transport activity in human normal and pancreatic cancer cells.

For transport studies, in-side out vesicles (IOVs) prepared from 20×106 cells was enriched for IOVs by wheat germ agglutinin affinity chromatography [26]. Transport activity was calculated from measurements of uptake of 14C-DOX (sp. activity, 8.5×104 cpm/nmol) and 3H-DNP-SG (sp. activity, 3.6×103 cpm/nmol) into the IOVs (20 µg/30 µl reaction mixture) in the absence or presence of 4 mM ATP as described [26]. Each transport experiment was done in triplicates in three separate experiments (n = 9).