Fig. 5

Arf6-mediated CD147 recycling promotes cell migration and invasion. a Transwell migration and invasion analysis. Equal numbers of Arf6-perturbed liver cancer cells were seeded into the upper chamber coated without or with Matrigel. Twenty-four hours later, cells migrating or invading through the Transwell filters were crystal-violet-stained and counted under a microscope. Representative results from three independent experiments are shown. The number of migrated or invaded cells in untreated groups (WT) was set as 100% for comparison. d MMPs activity analysis. Secreted MMPs in the concentrated medium (CM) were measured by gelatin zymography. Representative results from three independent experiments are shown. The MMPs secreted from untreated groups (WT) was set as 100% for comparison. f Cytoskeleton reorganization analysis. Arf6-perturbed cells were reseeded in poly-D-lysine-coated chamber slides, serum-free starved, fixed, permeabilized, and stained with Rhodamine-phalloidin. Representative confocal images from three experiments are shown. Scale bar: 20 um. g Rac1 activation analysis. Arf6-perturbed cells were serum-starved, lysed, and incubated with CRIB beads. The activated Rac1 in pull-down precipitates were immunoblotted, and representative results from three independent experiments are shown. The Rac1 activation level in untreated groups (WT) was set as 100% for comparison. b, c, e, h Cell counts and protein bands were quantified by Image J software, and significant differences compared with NC-KD cells are shown: n = 3, ** P < 0.01, * P < 0.05