Fig. 4

Effects of ATRA on markers of hypoxia in MCF10DCIS cells. a Proliferation of MCF10DCIS cells cultured in the presence of 1 μM ATRA for 96 h at normoxia or hypoxia. The data show the number of viable cells relative to normoxia, taken as 1. b Representative fluorescence microscopy images of MCF10DCIS cells grown on glass dishes for 96 h at normoxia or hypoxia in the presence of 1 μM ATRA or DMSO (vehicle) and subjected to immunocytochemical analysis with the anti-HIF-1α antibody. Bar: 20 μm. On the right, fluorescence intensity of digitized images calculated by the ImageJ software. c Representative Western blot analysis with the indicated antibodies of total lysates from MCF10DCIS cells under the same experimental conditions. Immunoblots shown have been cropped to conserve space. On the right, levels of CAIX as deduced from the densitometry of immunochemical bands normalized with β-Tubulin, used as internal control for equivalence of loaded proteins. Error bars indicate ± SD from a triplicate experiment. #P < 0.05 with respect to normoxia