Fig. 5.

hCTR1 knockdown in A2780 cells. (A) cDDP-sensitive (A2780) cells were transiently transfected with siRNA duplexes against CTR1 or siRNA negative control in Opti-MEM medium. Cells were biotinylated, and the biotinylated protein was analyzed on Western blots. hCTR1 protein was detected using an anti-CTR1 antibody (rabbit). (B) Biotinylated hCTR1 protein was quantified using Quality One software (Bio-Rad), and the protein levels were normalized to the Na⁺,K⁺-ATPase α subunit. After transient transfection, cells were incubated with 5 µM copper for 1 hour (C) or 30 µM cDDP for 3 hours (D). In (C), Cu uptake is expressed as a percentage of control (control rate 129.01 pmol of Cu/mg of protein/h), and in (D), cDDP uptake is expressed as a percentage of control (control rate 92.05 pmol of Pt/mg of protein/h). *Statistical analysis was performed using Student's t test, and P < 0.05 was considered statically significant.