Effects of BMP-2 on proliferation, migration and spheroid formation. A. Cells were treated in complete media with indicated doses as indicated or with 50 ng/ml BMP-2 or left untreated for four days. Cells were counted every two days and media was changed every second day. Values are the mean +/- SEM of duplicate wells from at least two independent experiments. B. BMP-2 increases the motility of cells. Wounds were made on confluent monolayers of cells and then treated at indicated doses or with 50 ng/ml BMP-2. After 20 hrs (TOV-2223) or 40 hrs (TOV-112D and TOV-1946) the open wound surface area was quantified by digital images taken under a phase contrast microscope. Values are the mean +/- SEM of duplicates in at least two experiments and represent the percentage of total area covered by the cells in each image. C. Effect of BMP-2 on spheroid formation. Cells were cultured using a modification of the hanging droplet method. Cells were incubated in media with or without 50 ng/ml BMP-2. Spheroid formation was monitored after ten days. Arrows show scattered cells. All pictures were taken at a magnification of 100×. For all figures * p < 0.05, ** p < 0.10. (Student t-test).