Fig. 6

EE-15-one does not affect cadherin dependent cell–cell adhesion. a To show that cell spheroids aggregate through Ca²⁺-dependent cell–cell adhesion, BT-20 cells (2 × 10⁴) were seeded in 1% agarose coated wells and allowed to form for 4 days as described in the materials and methods. Spheroids were then exposed to water (control), 2 mM EGTA (Low Ca²⁺), or 2 mM EGTA and 4 mM CaCl₂ (high Ca²⁺). Light microscopy images were taken after 24 h. b BT-20 spheroids were exposed to EE-15-one (5 µM), ESE-15-one (1 μM), 500 nM paclitaxel as positive control or DMSO (vehicle control). c The area and perimeter of the spheroid were measured and used to determine volume. Day 4 was set as staring volume and at each point the average of 60 independent spheroids volumes are represented. Error bars indicate standard deviation. Light microscopy images were taken at day 4, 7 and 15 at 10× magnification. Scale bar = 100 µm