Fig. 4

Effects of the intracellular domain of epithin/PRSS14. a The wound healing migration of 427, 427(SP2bKD)-10, and -16 cells in PMA-stimulated or non-stimulated conditions was analyzed. Data are presented as the means of the recovered area (six fields per each cell type, n = 3). b Invasion of 427, 427(SP2bKD)-10 and -16 cells in the presence or absence of PMA was analyzed. Numbers of migrated cells to the other side of the chamber for 24 h were counted, normalized against that of 427 cells, and shown as fold changes (n = 3). c Representative bright-field images of 427, 427(EICD)-1, and -5 cells are shown. The percentage of scattered cells over the total number of cells (> 745 cells in 50 fields) in each cell type is represented as a bar graph. Scale bars, 50 μm. d The degrees of migration of 427, 427(EICD)-1, and -5 for 24 h in the wound healing assay are quantified (six fields per each cell type, n = 3) and shown as bar graphs. e Invasion of 427, 427(EICD)-1, and -5 cells through transwell pores covered with Matrigel were analyzed as in b (5 fields per each cell type, n = 3). Statistical analysis was performed using an unpaired two-tailed Student’s t test. f A diagram of the lung metastasis assay is depicted. g The GFP fluorescence of 4T1KD plus EICD-transduced 4T1KD cells (4T1KD/EICD:GFP) was analyzed by flow cytometry and is shown as a histogram. h Representative lungs from mice injected with PBS (left) or 4T1KD/EICD:GFP cells (middle, right) are shown. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Error bars indicate SEM in all panels. i The percentage of GFP-positive cells in the mixed cells before injection and the GFP-positive nodules in five lungs after injection are shown. Error bars indicate SD