Fig. 3

Roles of CCDC88A in cell migration and invasion. a. Western blotting of CCDC88A following transient transfection of S2-013 and PANC-1 cells with a single mixture containing four different siRNA oligonucleotides targeting CCDC88A (siCCDC88A) or negative scrambled control (Scr). Western blotting was performed using an anti-CCDC88A antibody. b, c. Oligonucleotides targeting CCDC88A or Scr were transiently transfected into S2-013 and PANC-1 cells. Migration (b) and two-chamber invasion assays (c) were performed. Migrating cells in four fields per group were scored. Data are derived from three independent experiments. Columns, mean; bars, SD. *p < 0.005 compared with Scr-transfected control (Student’s t-test). d, e. Western blots (d) and confocal immunofluorescence microscopic images (e). Mock control vector or a myc-tagged CCDC88A-rescue construct was transiently transfected into scrambled control-siRNA and CCDC88A-siRNA transfected S2-013 and PANC-1 cells; after 48 h, the cells were incubated on fibronectin for 5 h. Western blots probed with anti-myc and anti-CCDC88A antibodies are shown. Cells were stained with anti-myc antibody (green). Actin filaments were labeled with phalloidin (red). Arrows, myc-tagged CCDC88A localized in cell protrusions. Blue, DAPI staining. Bar, 10 μm. f, g. Mock control vector or a myc-tagged CCDC88A-rescue construct was transiently transfected into scrambled control-siRNA and CCDC88A-siRNA transfected S2-013 and PANC-1 cells; 48 h later, migration (f) and two-chamber invasion assays (g) were performed. Migrating cells in four fields per group were counted. Data are derived from three independent experiments. Columns, mean; bars, SD. *p < 0.005 compared with corresponding CCDC88A-siRNA transfected S2-013 cells that were transfected with mock vector (Student’s t-test)