Fig. 3.
Effects of DEL-22379 on cell spreading, adhesion, migration and invasion of thyroid-tumor cells. 8505c and OCUT2 cells carry BRAFV600E and CAL62 and HTH83 cells carry K-, and H-RAS mutations, respectively. A Confocal immunofluorescence showing F-actin (red), paxillin (green) and overlay plus DAPI (blue) nuclear staining in OCUT2 (left panels) and CAL62 (right panels) cells, in the presence of 10% FBS plus vehicle (DMSO), 1 µM DEL-22379 or 10 µM U0126 for 48 h: scale bar is included in the red images, 20 µm. Arrows point to cells with increased spreading and arrowheads to focal adhesion sites. B Cell adhesion assay showing the relative mean (SD) from at least 3 independent experiments of the number of adherent cells 30 (8505c), 45 (OCUT2 and CAL62) and 20 (HTH83) min after seeding in 10% FBS-medium containing vehicle (DMSO), 1 µM DEL-22379 (D1), 5 µM DEL-22379 (D5), 10 µM DEL-22379 (D10) or 10 µM U0126. Cells were pre-treated for 30 min with the different agents in suspension before seeding. C Matrigel invasion assay showing the relative mean (SD) from 3 independent experiments of the number of invasive cells, after 24 h in serum-free medium containing vehicle (DMSO), 1 µM DEL-22379 (D1), 2.5 µM DEL-22379 (D2.5) or 10 µM U0126. D Wound healing assay showing the relative mean (SD) from 3 independent experiments of occupied cell-free area after 20 h in the presence of 10% FBS plus vehicle (DMSO), 1 µM DEL-22379 (D1), 2.5 µM DEL-22379 (D2.5) or 10 µM U0126. B–D Statistical significance of differences elicited by the treatments compared with a vehicle was estimated using a two-tailed t-test. (*)p = 0.05–0.01, (**)p = 0.01–0.001, (***)p < 0.001