Figure 2.

Overexpression of ZNF518B in RKO cells shows the nuclear location of ZNF518B and an increase in cell migration, invasiveness and adhesion to type I collagen. The cells were transfected with pCMV3-ZNF518B, or with the empty plasmid. (A) RT-qPCR analysis of the overexpression of ZNF518B in RKO cells. (B) Immunocytochemical analysis; size bar, 20 μm. (C) Transwell migration assay. A photograph of representative plates is shown (left) and the counting values (six different areas) were averaged (right). (D) Effects of overexpression of ZNF518B on the invasiveness of RKO cells. The assay was carried out as in (C), but through a Matrigel layer. (E) Effects of overexpression of ZNF518B on the adhesion of RKO cells to type I collagen-coated plates. The total number of cells was counted as in (C). (F) Colony formation assays carried out in six-well plates as described under Materials and Methods. Two independent experiments (each one with three independent transfections) were done with RKO cells. The photograph of representative plates and the averaged quantification of the number of colonies in each of the three wells are given. Statistical analyses were carried out by Student’s t-test. ***, p < 0.001.