Figure 5. Knockdown of Dub3 inhibits migration, invasion and CSC-like characteristics in BLBC cells by downregulation of Snail1.

(a) Dub3 was knocked down by two different shRNA in MDA-MB231 and MDA-MB157 cells. Rescued Snail1 expression in these Dub3-knockdown clones were also performed. The expression of E-cadherin, Claudin-7, N-Cadherin, Vimentin, Dub3, and Snail1 was analysed by western blot. (b) IF images of EMT markers in MDA-MB231 cell lines described in (a). Scale bars, 20 μm. (c) Graphic representation of cell motility described in a analysed by a wound healing assay. Data are the percentage of migrating cells as the mean±s.e.m. of three separate experiments. (d) Graphic representation of cell invasion described in a. Data are the percentage of vector control values (mean±s.e.m. in three separate experiments in duplicates). (e) Cell trajectories of randomly selected cells described in a; each line indicates an individual cell's migration. (f) Graphic representation of primary and secondary tumorsphere-formation from cells described in a and are the mean±s.d. from three independent experiments (left panel). (g) Graphic representation of the CD44^high/CD24^low (top) and CD49f^high/CD24^low population from cells described in a was examined by FACS analysis and are the mean±s.e.m. from three independent experiments.