Fig. 2

Effects of hypoxia on motility and CD133 expression in MCF10DCIS cells. a Representative phase-contrast images of MCF10DCIS cells grown at normoxia or hypoxia for 96 h. The arrows indicate cells with an elongated shape, whose number is shown in the graphs on the right. Bar: 20 μm. b Representative immunochemical analysis performed with the indicated antibodies of lysates from MCF10DCIS cells grown at normoxia (N) or hypoxia (H) for 96 h. Immunoblots shown have been cropped to conserve space. On the right, levels of E-cadherin and Vimentin as deduced from the densitometry of immunochemical bands normalized with β-Tubulin, used as internal control for equivalence of loaded proteins. c XCELLigence-driven dynamic monitoring of migration and invasion through diluted Matrigel of MCF10DCIS cultured at normoxia or hypoxia for 96 h. The slope analysis, that describes the steepness, incline, gradient, and changing rate of the Cell Index curves over time, was shown. d Representative cytofluorimetrical evaluation of CD133 expression with a PE-conjugated antibody in MCF10DCIS cells cultured at normoxia or hypoxia for 96 h. The antigen expression was represented on a bi-parametric dot plot in which a gate was based on the fluorescence emitted after marking with non-specific antibody (Isotype control). The percentage of cells showing high cell surface levels of CD133 is indicated at the upper right of each panel, together with their mean fluorescence intensity (MFI). The mean of three separate experiments ±SD is shown on the right graphs

All the data are the mean of three separate experiments performed in triplicate ±SD. *P < 0.05