Fig. 3.

EMP1 induced increase in cell migration and invasion in LNCaP prostate cancer cells. a Several LNCaP clones stably expressing FLAG-EMP1 were verified by western blotting. b Proliferation of LNCaP cells with very low FLAG-EMP1 expression (clone #2) and high expression (clone #17) was assessed by cell counting at the indicated time points. c, e Representative results of the Boyden chamber and invasion assays. Cells migrated to the lower chamber were visualized by modified Giemsa staining. d, f Summary graph of migrated LNCaP cells in the Boyden chamber and invasion assays. *p < 0.05 and **p < 0.01 vs. LNCaP #2. g Collective cell migration assay. Confluent cell layers of each LNCaP cell clone were scratched, and cell migration was observed for 12 h by time-lapse light microscopy. Dotted green lines indicate the cell front immediately after scratching. Arrowheads indicate cells distant from the cell clump through particularly higher cell migration. Scale bars, 200 μm in c and e, and 100 μm in g