Cell migration, apoptosis and proliferation
Cell proliferation was performed by seeding NPC cells into 6-well plates
at the density of 1,000 cells/well, culturing for 7 days in the presence
or absence of different concentrations of IDX with/without cisplatin,
fixing in 4 % paraformaldehyde and staining with 1 % crystal violet.
For the migration assay, NPC cell lines were seeded on the upper
chambers of 24-well Transwell inserts (BD Biosciences) and allowed to
migrate with/without combined drugs in the bottom well. Migrated cells
were fixed and stained as described above. For the apoptosis assay, the
apoptotic cells were quantified (percentage) using an Annexin
V-FITC/propidium iodide (PI) apoptosis detection kit (BD Biosciences).
Cells were seeded at a density of 2 x 105 cells per
well in 6-well plates. After treatment, cells were harvested and
Annexin-V-FITC/PI labeling was performed according to the manufacturer’s
instructions. The stained cells were analyzed on a NovoCyte Quanteon
flow cytometer (Acea Biosciences).