FIGURE 6.
Effect of SK-1 overexpression on HPR sensitivity in A2780 cells. A2780 cells were transfected with the empty expression vector (mock transfected) or with pcDNA3-hSK1WT plasmid (SK1-overexpressing cells). A, Western blot analysis of FLAG expression in A2780 transfectants. Equal amount of proteins of A2780, mock, and SK1-overexpressing cells were loaded onto SDS-PAGE and then blotted onto a polyvinylidene difluoride membrane, and the overexpressed SK1 tagged with FLAG M2 epitope was detected by anti-FLAG M2 antibody. B, 12 h after seeding, A2780 and A2780 transfectants (one mock and 3 SK1-overexpressing clones, 1, 2, and 3) were treated with 1 μm HPR. After 48 h, the total cell number was determined by counting trypsinized cells using a Bürker counting chamber, and the number of dead cells was evaluated by the Trypan blue exclusion test. Data are presented as the percentage of the total counted cells and are the means ± S.D. of three different experiments. *, p < 0.01 versus A2780 cells.