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. 2019 Oct 4;10:682. doi: 10.3389/fendo.2019.00682

Figure 2.

Figure 2

24-well plate with a cell-repellent surface method. BON1 cells were seeded into a 24 well plate with a repellent surface, mixed overnight at 80 rpm. (A) Spheroids were treated with increasing Sunitinib concentrations and pictures were taken at day 4, 7, and 10 after seeding with a Zeiss Axiovert 200/M-based phase-contrast microscope (5 × objective). (B) Perimeter analysis of spheroids was performed at day 4, 7, and 10. Gray column: perimeter analysis at Day 4, before treatments. White columns: perimeter analysis at Day 7 and 10 under indicated treatments. The analysis was performed using Image J software and measurements were performed evaluating three independent experiments in two replicates. **P < 0.01 vs. vehicle cells at Day 10. § = 5 μM measurement was not detectable for technical reasons, as indicated in the results section. (C) Immunohistochemical expression of Caspase 3 in spheroids treated with different Sunitinib concentrations. Spheroids were fixed at day 10 and pictures were taken with a Zeiss Axiovert 200/M-based phase-contrast microscope. Pictures provide an overview of the entire spheroid stained with eosin and Caspase 3 antibody.