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. 2014 Mar 3;111(11):4049–4054. doi: 10.1073/pnas.1321562111

Fig. 1.

Fig. 1.

CmABCB1 is an ABC multidrug transporter. (A) Dendrogram of ABC transporters from C. merolae and human P-gp (ABCB1). Among 32 ABC proteins from C. merolae, CMD148C is the most similar to human P-gp and was therefore designated as CmABCB1. This dendrogram is based on multiple sequence alignments of the full amino acid sequences of ABC transporters. Alignments were performed using with ClustalW and visualized using Drawtree. (B) Drug-susceptibility assay in S. cerevisiae AD1-8u− cells. AD1-8u− cells expressing WT CmABCB1 (●) were grown in various concentrations of drugs. For each drug assayed, mock-transfected AD1-8u− cells (○) were also grown as controls. Data are means ± SD (n = 5). (C) Drug-susceptibility assay in HEK293 cells. Mock-transfected HEK293 (○) and CmABCB1-expressing HEK293 cells (●) were incubated with paclitaxel or vinblastine, and viability was measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Data are means ± SD (n = 3). (D) Effect of drugs on CmABCB1 ATPase activity. The ATPase activity was measured in the presence or absence of 50 µM of the indicated drugs with 5 mM ATP at 37 °C. Data are means ± SD (n = 2). (E) Drug concentration dependence of CmABCB1 ATPase activity. The ATPase activity was measured as a function of verapamil or rhodamine 6G concentration with 5 mM ATP at 37 °C. Data are means ± SD (n = 3). The solid line is a fit of the equation to the data as described in SI Materials and Methods (Eq. S2).