Immunoblot detection of MRP7 and P-gp and the effect of imatinib or nilotinib on MRP7 expression.

<p>(A) Expression of MRP7 in HEK293-pcDNA3.1 (lane 1) and MRP7-transfected cells (lane 2). (B) Expression of P-gp in HEK293-pcDNA3.1 (lane 1), HEK-MRP7-2 (lane 2), KB-3-1 (lane 3) and KB-C2 cells (lane 4). (C) Effect of 5 µM of imatinib on the expression level of MRP7 (HEK-MRP7-2) for 36 and 72 h, respectively. (D) Effect of 5 µM of nilotinib on the expression level of MRP7 (HEK-MRP7-2) for 36 and 72 h, respectively. Equal amounts (40 µg protein) of total cell lysate were used for each sample. The nitrocellulose membranes were immunoblotted with primary antibody against MRP7 or actin at 1∶500 dilution, or P-gp at 1∶500 dilution at 4°C overnight, and then incubated with HRP-conjugated secondary antibody at 1∶1000 dilutions at room temperature for 3 h.