Fig. 4

Overexpression of CDH4 improved the proliferation, stemness and motility of OSCC. CDH4 expression was effectively increased by pcDNA3.1-FLAG-CDH4 plasmid as measured by western blot A. The growth of HN30 cells was measured by CCK-8 assay B. The colony formation assay showed that the proliferation of HN30 cells was increased after CDH4 up-regulation C. The number of colonies was counted D. The picture shows the cell sphere formation in different groups under a 40X microscope E, and the number of spheres was increased in the CDH4-overexpression group compared with the vector group F. For the wound-heal assay, the representative images show the distances of cells passing through the scratch within a period of time for HN30 cells G under a 40X microscope field of view. The mobility rate of cells was measured for HN30 cells H. In the transwell migration assay, representative images show the cells transfected that passed through the upper chamber of the transwell without (upper panel) Matrigel coating for HN30 cells I. The number of cells that passed through the filters was counted for HN30 cells J. In the transwell invasion assay, representative images show the cells transfected that passed through the upper chamber of the transwell with (upper panel) Matrigel coating for HN30 cells K. The number of cells that passed through the filters was counted for HN30 cells L. The numbers of migrating cells were counted from at least five randomly selected fields of view under a microscope at 100X magnification. These experiments were repeated three times. (* indicates P < 0.05, **indicates P < 0.01, ***indicates P < 0.001, ****indicates P < 0.0001, Values are presented as means ± standard error.)