Figure 1.

Irradiation-Induced Changes in the Vasculature of BM and Spleen

(A) Schematic representation of protocol for lethal irradiation analysis.

(B) Tile scan overview images of Emcn-stained vessels in adult femur after irradiation.

(C) Emcn, CD31, and collagen IV immunostaining of control and irradiated BM, as indicated. Arrows mark Emcn⁺ CD31⁺ vessels in middle panel and collagen IV⁺ reticular fiber on the right.

(D) Quantification of Emcn⁺ area in imaging field (n = 6 per group).

(E) Morphology of individual ECs (arrows) in control and irradiated bones of Cdh5-CreER R26-mTmG mice. Low dosage of tamoxifen was injected 6 days after irradiation.

(F) Quantification of area, perimeter, and shape factor from control (n = 147 from 3 mice) and 9 Gy (n = 140 from 4 mice) single ECs. Shape factor is a numerical indication of how similar a 2D shape is to a perfect circle, which has a shape factor of 1.

(G) Nuclear GFP⁺ (nGFP⁺) ECs in control and irradiated Cdh5-mTnG diaphysis. Graph shows quantitation of GFP⁺ cells (n = 6 in each group).

(H) FACS plot of GFP⁺ cells from control and irradiated Cdh5-mTnG mice. Graph show frequency of GFP⁺ cells (n = 20 in each group).

(I) Tile scan overview images and selected maximum intensity projections of spleen vessels in Cdh5-mTnG mice. Quantification of GFP⁺ nuclei in each image field (Ctrl n = 6; 9 Gy n = 4) is shown.

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

See also Figure S1.