These
assays were performed as previously described.5 (link),40 (link),41 (link) Briefly, cells were seeded in six-well plates
(100 cells/well for PANC-1 and 200 cells/well for BxPC-3) and cultured
for 24 h before addition of PPIs or DMSO vehicle. The cells were continuously
cultured in the presence of PPIs or DMSO for 10–14 days followed
by staining with crystal violet and counting manually.
For the
apoptosis assay, BxPC-3 cells were seeded in 12-well plates (18 000
cells/well) and cultured for 24 h before treatment with lansoprazole
or DMSO control for 72 h. The cells were then harvested and subjected
to analysis using the cell death detection ELISA assay kit (Roche)
according to the manufacturer’s instructions. For detection
of cleaved PARP, BxPC-3 cells were seeded in six-well plates (200 000
cells/well) and cultured for 24 h before treatment with lansoprazole
or DMSO control for 24 h. Cells were then collected and subjected
to Western blot analysis of cleaved PARP using an antibody specific
to the cleaved PARP (Cell Signaling).
assays were performed as previously described.5 (link),40 (link),41 (link) Briefly, cells were seeded in six-well plates
(100 cells/well for PANC-1 and 200 cells/well for BxPC-3) and cultured
for 24 h before addition of PPIs or DMSO vehicle. The cells were continuously
cultured in the presence of PPIs or DMSO for 10–14 days followed
by staining with crystal violet and counting manually.
For the
apoptosis assay, BxPC-3 cells were seeded in 12-well plates (18 000
cells/well) and cultured for 24 h before treatment with lansoprazole
or DMSO control for 72 h. The cells were then harvested and subjected
to analysis using the cell death detection ELISA assay kit (Roche)
according to the manufacturer’s instructions. For detection
of cleaved PARP, BxPC-3 cells were seeded in six-well plates (200 000
cells/well) and cultured for 24 h before treatment with lansoprazole
or DMSO control for 24 h. Cells were then collected and subjected
to Western blot analysis of cleaved PARP using an antibody specific
to the cleaved PARP (Cell Signaling).
