Figure 3.

ULA plate method. BON1 cells were seeded into an ultra low attachment 96 well plate and spheroids were obtained by centrifugation. (A) Spheroids were treated with increasing Sunitinib concentrations and pictures were taken with EVOS FL Cell imaging System (10 × objective) at Day 3 and 7 after seeding. (B) Perimeter analysis of spheroids was performed at Day 3 and 7. Gray column: perimeter analysis at Day 3, before treatments. White columns: perimeter analysis at Day 7 under indicated treatments. The analysis was performed using Image J software and measurements were performed from three independent experiments in two replicates. *P < 0.05 vs. vehicle cells. (C) Cell metabolic activity was measured as absorbance in three independent experiments with six replicates each, and it is expressed as the mean ± S.E.M. **P < 0.01 vs. vehicle cells.