Figure 4.

KRT16 knockdown in MDA-MB-468 impaired cell migration (A) mRNA expression level was verified after KRT16 knockdown compared to the non-target (siRNA control). (B) Immunoblot to verify the expression levels of CDH1 and VIM in KRT16 depleted cells. The relative ratio of protein expression of (C) K16; (D) CDH1; and (E) VIM. (F) Relative mRNA expression of EMT/MET-specific genes was investigated after KRT16 depletion; mRNA expression values were normalized to GAPDH and subsequently displayed relative to gene expression as a fold change 2^{−(∆∆CT)}. (G) Microscopy images of migrated cells by wound-healing assay were performed on KRT16 depleted cells (KRT16 siRNA) compared to control cells (siRNA control) at different time points of 0 h, 24 h, 72 h, and 120 h; n = 2, scale bar represents 100 μm. (H) In migration, linear regression analysis was performed for the velocity slope of KRT16-siRNA transfected cells and untreated cells (control). (I) Cell proliferation rates of KRT16-siRNA transfected and siRNA control cells were determined using Trypan Blue Dye, and the cells were counted by a Vi-CELL Counter device, the data are expressed as the mean ± SD.; n = 3. The p-values were calculated by the Wilcoxon rank-sum test (p < 0.05).