Extended Data Fig. 6: Intravital microscopy of virgin and involuting mammary glands.
From: Tissue-resident ductal macrophages survey the mammary epithelium and facilitate tissue remodelling
a, Photo of a mouse prepared for intravital imaging (left) and the exposed mammary gland after intraductal injection of fluorescent beads (right). b, Isolation of DM MHCII signal in 3D-IVM images from Elf5-GFP mice with labelling by anti-MHCII AF647 antibody (see Methods, 6 experiments). Left: raw GFP (magenta) and MHCII (cyan) signal. Middle: addition of GFP surface rendering (white). Right: GFP and masked duct-adjacent DM MHCII signal (yellow). Scale bar, 30 µm. c, Time-points from a 3D-IVM movie (Supplementary Video 3) in which precise laser damage was induced in the epithelium of a Cx3cr1GFP/+ mouse at 9 weeks. Images representative of 3 mice. Time hrs:mins. Scale bar, 20 µm. d, 3D-IVM of Elf5-GFP mammary tissue at 3 days involution (Supplementary Video 5) showing GFP (magenta) and anti-MHCII AF647 antibody (yellow). Left: overview. Right: enlarged 3D projections at time-points throughout phagocytosis and the outlined volume viewed from the side. Arrows indicate GFP+ cells within DMs. Images acquired every 5 mins. Images representative of 3 mice. Scale bars, 100 µm (left), 20 µm (enlargements).
