ZIP4 silencing inhibited migration and invasion capabilities of C666-1 cells.a Relative protein and mRNA level of ZIP4 were detected in stable LVRH, Sh-ZIP4, Control, and ZIP4 cell lines. b The cell migration rate between LVRH C666-1 and Sh-ZIP4 C666-1 cells was compared by the wound-healing assay. Microscopic observation was recorded at 0 and 24 h after scratching the cell layer (Scale bar = 50 μm). c The invasive and migration properties of the cells were analyzed by using a matrigel-coated Boyden chamber. Migrated cells were plotted as the average number of cells per field from three different experiments (Scale bar = 50 μm). d The expression levels of EMT markers, E-cadherin (E-cad), vimentin (Vim), and FSP-1 were examined by immunofluorescence staining (Scale bar = 75 μm). e E-cad, vimentin (Vim), and FSP-1 expression were examined in control and ZIP4 C666-1 cells by western blot. The experiments were performed in triplicate. The data are represented as means ± SD from three independent experiments. *P < 0.05
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