Fig. 2.

Myoepithelial cells (MEPs) in mammary architecture and microenvironment engineering (MAME) cocultures facilitate organization of MCF-10A acinar structures and reduce size and dysplastic phenotype of ductal carcinoma in situ (DCIS) structures. Nontransformed breast epithelial cells (MCF-10A) and MCF10.DCIS cells (DCIS) were grown in MAME cultures in the absence and presence of N1ME cells (MEPs) and imaged live at 6 and 21 days. Red and green represent phalloidin staining of the actin cytoskeleton and immunostaining for human laminin-332, respectively. a 3D reconstructions of Z-stack images captured at 6 (top rows) and 21 (bottom rows) days in MAME culture and generated with Volocity software. One grid unit = 23 μm. Inset in top row, left column, is a representative en face view of MCF-10A plus MEPs. b Confocal slices at the equatorial plane of the structures captured at 6 (top rows) and 21 (bottom rows) days in MAME cultures. Scale bar = 50 μm. Inset in top row, left column, is a representative image of MCF-10A cells grown alone in MAME culture at day 6. Images are representative of at least three independent experiments. Acinar lumens (white arrows) and tubular morphology (white bracket) are indicated. Immunostaining for laminin-332 and videos of MCF-10A 3D acinar structures are shown in Additional file 1: Figure S1, Additional file 2: Video S1, Additional file 3: Video S2, and Additional file 4: Figure S2