Figure 4: Temporal control of AURKA expression is important for the generation of binucleated alveolar cells.
From: Essential role for a novel population of binucleated mammary epithelial cells in lactation
(a) Whole-mount 3D confocal images of ductal portions and optical sections from the enlargements for Elf5-rtTA/TetO-cre/AURKAf/f gland (bottom) compared with control TetO-cre/AURKAf/f mammary gland (top). The whole-mounts were labelled for DAPI (white), GFP (green), milk (red) and F-actin (blue). The animals were given DOX food from 16.5 dP until imaging analysis at 5 dL (n=4 mice). Scale bars: 100 μm (whole-mounts); and 20 μm (optical sections). (b) Bar graph of FACS data showing percentage of cells containing 2N or 4N DNA content in the luminal, basal and stromal cell compartments from Elf5-rtTA/TetO-cre/AURKAf/f (AURKAΔ/Δ) and control AURKAf/f mice at 2 dL, after DOX induction from 16.5 dP. Error bars represent mean±s.e.m. (n=3 mice). **P<0.01. (c) Body weight of pups nursed by Elf5-rtTA/TetO-cre/AURKAf/f dams versus littermate control TetO-cre/AURKAf/f dams. DOX food was given from 16.5 dP to 14 dL (six pups per dam; n= 3 dams). P<0.0001. (d) Body weight of pups nursed by Elf5-rtTA/TetO-cre/AURKAf/f dams versus control dams. DOX was given from 4 to 14 dL (six pups per dam; n=2 dams). Error bars represent mean±s.e.m. (no significant difference). (e) Representative whole-mount 3D confocal image of a ductal portion from Elf5-rtTA/TetO-cre/AURKAf/f mice. DOX food was given to mice from 4 dL until collection at 10 dL. The whole-mount was labelled for DAPI (white), E-cadherin/GFP (green), K5 (red) and F-actin (blue) to highlight the architecture of the gland (n=2 mice). Middle and right panels show optical sections from the selected area in the left panel. Most cells remain binucleated. Scale bars: 200 μm (whole-mount); and 30 μm (optical sections).
