Fgd5^{ZsGreen•ZsGreen/+} reporter expression marks all HSC subtypes and also endothelial cells in the BM. (A) BM ZsGreen⁺ (green histograms) and ZsGreen⁻ (gray histograms) cells of Fgd5^{ZsGreen•ZsGreen/+} mice co-stained and gated through lineage (Ter119, Mac-1, Gr-1, B220, CD3, CD4, CD8, and IL7Rα), c-Kit, Sca1, CD48, and CD150 (n = 5 or more). (B) Sub-fractionation of LKSFlk2⁻CD34^−/low into CD150^hi (red gates), CD150^lo (green gates), and CD150^neg (blue gates) HSC subtypes from young (4 mo) and mid-aged (14 mo) Fgd5^+/+ and Fgd5^{ZsGreen•ZsGreen/+} mice. (C) Histograms showing ZsGreen expression of each of the indicated HSC subtypes from the mice shown in B. MFI of each of the indicated HSC subtypes is shown (bottom). A minimum of n = 3 mice for each stained were analyzed with the exception of 14-mo-old mouse (n = 1). (D) Adult femoral BM cavity microvascular network based on 3D reconstruction of a series of confocal images showing arterial and sinusoidal microvessels (stained with Laminin in red) and Fgd5^{Zsgreen•ZsGreen/+} reporter expression (green). Projection depth 54 µm. Grid squares (white box) are 51.3 µm × 51.3 µm (left) and 42.9 µm × 42.9 µm (right).