Figure 3.

Diphtheria Toxin-Mediated Hematopoietic Cell Ablation Mimics Irradiation-Induced Vascular Defects

(A) FACS plot of LSK cells 4 days after diphtheria toxin injection in DTR^iΔHC and control mice.

(B) Tile scan overview images of Emcn-stained vessels in DTA-treated control and DTR^iΔHC long bone.

(C) High-magnification images showing Emcn, CD31, and collagen IV immunostaining in DTR^iΔHC and control femur.

(D) Tile scan images showing extravasation of fluorescent Dextran in DTR^iΔHC and control long bone.

(E) Cdh5-mTnG–controlled GFP⁺ EC nuclei and Emcn staining in DTR^iΔHC and control diaphysis.

(F) Quantification of LSK cell number (Ctrl = 4, DTR^iΔHC= 5), Emcn⁺ area (Ctrl = 7, DTR^iΔHC= 7), Dextran⁺ area (Ctrl = 4, DTR^iΔHC= 3), and GFP⁺ cells per image (Ctrl = 7, DTR^iΔHC= 7, frequency of GFP⁺ cells by FACS (Ctrl = 11, DTR^iΔHC= 8).

(G) Representative images and quantification of Emcn⁺ area at 16 days after irradiation and transplantation of 4 × 10⁵ Lin^– or Lin⁺ cells (n = 4 in each group)

(H) Emcn, GFP⁺ EC nuclei, CD31, and collagen IV at 16 days after irradiation and transplantation of 2 × 10⁴ Lin^– or LSK cells. Quantification of Emcn area (LSK = 10, Lin^– = 9) and GFP⁺ EC nuclei in each image field (LSK = 9, Lin^– = 8).

(I) Tile scan overview images showing Dextran extravasation at 16 days after irradiation and transplantation of 2 × 10⁴ Lin^– or LSK cells. Quantification of Dextran⁺ area (LSK = 5, Lin^– = 5).

Error bars, mean ± SEM. p values, two-tailed unpaired Student’s t test. See also Figure S3.