Fig. 5

BMP1 inhibitor UK 383367 inhibits mobility of gastric cancer cell line AGS and MKN45. (a) Relative BMP1 expression of AGS, MKN28, and MKN45 cells was determined by qRT-PCR and immunoblotting. (b) MKN28 cells were seeded into a culture insert for 24 h, followed by culturing with 2% serum plus UK 383367 for another 24 h. After removal of the culture insert, images were taken 0 and 8 h using an Olympus FV1000 laser confocal scanning microscope equipped with differential interference contrast system. ImageJ was used to measure the distance covered by migrating cells, and statistical analysis was performed using Student’s t-test. (c) AGS cells was treated with 400 nM, 800 nM, or 1600 nM UK 3367, and the mobility of treated cells was assessed by wound healing assay as described in (b). (d) The mobility of MKN45 was assayed as described above. (e) MKN45 cells were seeded into the upper chamber of a transwell with or without UK 383367 and cultured for 24 h. Serum was then removed from the upper chamber, and the cells were allowed to move for 24 h. Cells were then fixed, stained with DAPI, and imaged using an Olympus FV1000 laser confocal scanning microscope. The percentage of the cells migrating to the lower chamber was calculated. Two independent experiments were carried out