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. 2014 May 7;2014:382653. doi: 10.1155/2014/382653

Figure 4.

Figure 4

Expression of autophagic proteins (Atg5, Atg3, LC3, and beclin-1) and phosphorylated Bcl-2 (Ser70) in BME-UV1 cells forming acinar structures on Matrigel. Representative images of Western blot analysis of Atg5 (a), Atg3 (b), LC3 (c), beclin-1 (d), and phosphorylated Bcl-2 (Ser70) (f) in bovine MECs grown in 3D culture for 3, 6, 9, and 14 days in differentiation medium (control), enriched with 17β-estradiol (E2, 1 nM), progesterone (P4, 5 ng/mL), or both (E + P); expression of β-actin was used as a loading control; graphs below the images show the 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. (e) Confocal images of immunofluorescence staining of beclin-1 (green fluorescence) in cells grown in 3D culture for 3 or 9 days in control conditions: DNA was counterstained with 7AAD (red fluorescence) and white arrows indicate the nuclear localization of beclin-1.