Fig. 2

RAB4A knockdown changes the expression of well-known EMT genes in MDA-MB-231 breast cancer cells, in the opposite direction as expected for EMT. The complete profile of the 88 EMT genes whose mRNA levels were determined is presented in Additional file 1: Fig. S2. A The table presents the top five up- and down-regulated EMT-related genes in RAB4A knockdown cells in comparison to the control cells; the changes in expression are in the opposite directions as predicted for EMT (last column) [40,41,42]. B qPCR validation of the gene expression of top EMT markers in response to RAB4A knockdown. Data is presented as mean ± SEM (n ≥ 5). “*” represents p < 0.05 relative to the expression in control cells, which is set as baseline. C Fluorescent microscopic analysis of CDH1 (E-cadherin) expression in MDA-MB-231 cells transfected with control or two different RAB4A KD sequences as in (A) and (B). DAPI was used to stain the nucleus. Scale bar = 100 µM. D Invasion assay of control and RAB4A knockdown MDA-MB-231 cells. Plates were visualized after 24 h in the matrigel invasion culture. Scale bar = 100 µm