Functional Interactions between 17β-Estradiol and Progesterone Regulate Autophagy during Acini Formation by Bovine Mammary Epithelial Cells in 3D Cultures
Figure 2
Cleaved caspase-3 expression in BME-UV1 cells cultured on Matrigel in differentiation medium (control), enriched with 17β-estradiol (E2, 1 nM), progesterone (P4, 5 ng/mL), or both (E2 + P4) for 3, 6, 9, 12, or 14 days; (a) images present immunofluorescence staining of cleaved caspase-3 (green fluorescence) and DNA counterstained with 7AAD (red fluorescence) and graph beside represents quantitative analysis of the intensity of green fluorescence of cleaved caspase-3 immunostaining, presented as the ratio of integrated optical density (IOD) of caspase-3 to IOD of nuclei in each acinus analysed; (b) Western blot analysis of the levels of cleaved caspase-3 in cell lysates: expression of β-actin was used as a loading control; graph beside the image represents the obtained results of densitometric analysis, in which IOD of each band was measured, and the values were normalized to IOD of β-actin; the IOD results are presented as means ± SEM from at least three separate experiments; statistically significant differences () in comparison with control conditions were marked with .
