Quantifying ABCB1 Expression in TAK-243-Treated KB-C2 Cells

ABCB1-overexpressing KB-C2 cells were incubated with TAK-243 for up to 10 days. The total soluble protein lysates were prepared, subjected to SDS-PAGE, and transferred to PVDF membranes [44 (link)]. To quantify the expression level of ABCB1, membranes were probed with anti-ABCB1 monoclonal antibody (C219) (MA1-26528, Thermo Fisher Scientific Inc., Waltham, MA, USA). The loading control GAPDH was probed by anti-GAPDH monoclonal antibody (GA1R) (MA5-15738-D680, Thermo Fisher Scientific Inc., Waltham, MA, USA). Mouse IgG HRP conjugates (#7076, Cell Signaling Technology Inc., Danvers, MA, USA) were used as secondary antibody. To visualize the bands, ECL detection reagents were used (Thermo Fisher Scientific Inc., Waltham, MA, USA). The protein band densitometry was measured using ImageJ software (NIH, MD, USA).