Fig. 5

TGF-β1 induction increases the CD44+/CD24- subpopulation by coordinating gene expressions for CSC markers, EMT markers, and GSN in MDA-MB231 cells. a Fluorescence-activated cell sorting (FACS) analysis to measure CD44 and CD24 expression for MDA-MB231 cells with (right) or without (middle) additions of 2 ng/ml TGF-β1 for 3 days. CD44 and CD24 were detected by a combination of fluorochrome conjugated monoclonal antibodies against human CD44 (APC) and CD24 (PE), respectively. Mock (left) used as unstained control MDA-MB231 cells without TGF-β1. Data acquisition as shown below for each plot analysis. Real-time quantitative PCR (qPCR) analysis showing the increased gene expression for markers, including (b) stem cell pluripotency (i.e. Oct4, Sox2 and Nanog), (c) mesenchymal cell markers such as N-cadherin, vimentin, and E-cadherin, and for (d) GSN in post-sort CD44+/CD24- sub-population of MDA-MB231 cells with additions of 2 ng/ml TGF-β1 for 3 days. In b, c, and d, each value is the mean ± SEM (n = 6). *indicates significant difference compared to mock control, and # symbolizes a significant difference compared to post-sort CD44+/CD24- sub-population of MDA-MB231 cells without TGF-β1 treatment