Fig. 2

Influences of cortisol treatment on 3D growth of mammary epithelial cells for 14 days identified by immunofluorescence. Cells were treated with cortisol 0.7 μM or vehicle (methanol) from day 5 after seeding until day 14. a Upper panels: immunodetection in primary epithelial and myoepithelial cells of K14 (myoepithelial cells), K19 (epithelial cells) and Hoechst dye was used as nuclei counterstaining. Scale bar = 50 μm. Bottom panels: quantification of morphometric analysis in the control group and cortisol-treated group of the number of acini formed and quantification of disrupted acini per total number of acini. b Upper panels: immunofluorescence of laminin (BM), cytokeratin14 (myoepithelial cells) and Hoechst dye to counterstain nuclei. Scale bar = 100 μm. Bottom panels: quantification by Image J software of laminin intensity after cortisol 0.7 μM or vehicle treatment. c Immunofluorescence in MCF10DCIS 3D growth of laminin (basement membrane), Muc1 (epithelial cells) and Hoechst dye to stain the nuclei under control (vehicle), cortisol 0.7 μM, RU486 0.5 μM and cortisol 0.7 μM + RU486 0.5 μM. Arrows indicate rupture points of the acini. d Morphometric quantification of disrupted acini and acinar fusion and intensity of laminin determined by the integrated density parameter of Image J software. Scale bar = 50 μm. All experiments were carried out in triplicate. Statistical analysis was performed using the Mann-Whitney test or one-way analysis of variance followed by Tukey post-hoc test