Figure 5

ATP-dependent transport of TCA (○), GCA (▲) and CA (□) in membrane vesicles (7.5 μg protein) expressing human, dog and cat BSEP/Bsep. Vesicles were incubated in 10 mMTris buffer (pH 7.4) containing 250 mM Sucrose, 10 mM MgCl₂ and 4 mM ATP or AMP, at 37°C for 5 min (TCA) or 7.5 min (CA and GCA). ATP-dependent transport was calculated by subtracting transport in presence of AMP from that in presence of ATP. Curves were fitted by using GraphPad Prism 6.01 software (San Diego, California, USA). Measurements were performed in duplicate in three independent experiments and are given in pmol/mg protein/min. Each value in the graph is the mean ± SD of three independent experiments.