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. 2020 Aug 14;12(8):2275. doi: 10.3390/cancers12082275

Figure 2.

Figure 2

Evidence of increasing Annexin A8 (ANXA8) upregulation in isogenic MCF10A cell lines. (A) Immunofluorescence of MCF10A-Ctrl, MCF10A-1H, and MCF10A.com cell lines grown in two-dimensional (2D) culture stained with both 4′,6-diamidino-2-phenylindole (DAPI) (blue) and ANXA8 antibody (green) (scale bar 30 µm). (B) Western blotting (WB, top) and ANXA8 quantitative protein expression (bottom) in the three cell lines show an increasing ANXA8 level. (C) MCF10A-Ctrl, MCF10A-1H, and MCF10A.com cells grown in three-dimensional (3D) basement membrane culture (Matrigel) for 12 days were fixed and stained for DAPI and ANXA8 and imaged by confocal microscopy. MCF10A-Ctrl cells formed 3D acinar structures with a lumen lined by ANXA8-positive cells, while MCF10A-1H cells stably expressing the RAS oncogene developed morphologically aberrant acinar structures with ANXA8-positive cells in the luminal space. Comedocarcinoma MCF10A.com cells formed 3D ANXA8-positive “grape-like” morphologically aberrant structures (scale bar 30 µm).