Figure 4
From: The Ras-related gene ERAS is involved in human and murine breast cancer
Induction of EMT and stem cell-like features by ERAS expression in mammary gland cells. (a) Contour plots of flow cytometry analysis showing EpCAM and CD49f (integrin α6) immunostaining for each cell type. (b) Quantification of data shown in a. MCF10A control cells present both EpCAM+/CD49f+high and EpCAM−/CD49f+low subpopulations, showing mainly an epithelial phenotype, whereas almost all MCF10A-ERAS cells were EpCAM−/CD49flow (p < 0.0001). Data represent the means ± SD from three independent experiments. (c) Gene-expression levels of transcriptional factors measured by RT-qPCR. Values represent the mean ± SD from three different experiments and were normalized by TBP expression and represented in respect to control cells. (d) Western blot analysis showing the switch between E-cadherin and N-cadherin in mammary gland cells expressing ERAS. Uncropped blots are presented in Supplementary Fig. S6. (e,f) Immunofluorescences of HA-tag, E-cadherin and N-cadherin in co-culture of MCF10A control and MCF10A-ERAS cells. Bar, 20 μm (g) Representative contour plots of flow cytometry showing CD44 and CD24 immunostaining for each cell type. (h) Quantification of data shown in G. CD44high/CD24− subpopulation is 1–2% in control cells, however this subpopulation is majority (~80%) in MCF10A-ERAS cells (p < 0.0001). Data represent the means ± SD from three independent experiments.
