Fig. 3: Hypervascularization and perivascular hypercellularity in male grafts placed within female hosts.

a Representative image of blood vessels within GFP⁺ graft. The vasculature is visualized by CD31 immunoreactivity on vascular endothelial cells; outlines of blood vessels are drawn with dotted lines. b High-magnification images of grafts showing blood vessels outlined with dotted lines. c Quantification of the total vascular area within grafts. MM (n = 9 mice); MX (n = 4 mice); MF (n = 9 mice); FF (n = 5 mice); FX (n = 8 mice); FM (n = 7 mice). MM vs. FM: P < 0.0001; MX vs. FM: P = 0.0008; MF vs. FM: P < 0.0001; FF vs. FM: P < 0.0001; FX vs. FM: P < 0.0001. d Quantification of vascular diameter within grafts. MM (n = 6 mice); MX (n = 4 mice); MF (n = 7 mice); FF (n = 5 mice); FX (n = 8 mice); FM (n = 7 mice). MX vs. FM: P = 0.0030; FF vs. FM: P = 0.0228; FX vs. FM: P = 0.0003. e Representative images of grafts labeled with nuclear marker DAPI. Hypercellularity is evident by increased numbers of DAPI⁺ nuclei around blood vessels in the female host/male graft group (asterisks). f Process of drawing perivascular ROIs. First, ROIs were drawn for blood vessels (left), then ROIs were enlarged by 50 μm (middle), and the number of DAPI + nuclei were quantified within enlarged perivascular ROIs (right). g Quantification of DAPI⁺ cell density within the 50 μm perivascular zone surrounding blood vessels in grafts. MM (n = 9 mice); MX (n = 4 mice); MF (n = 9 mice); FF (n = 5 mice); FX (n = 6 mice); FM (n = 9 mice). MM vs. FM: P = 0.0058; MX vs. FM: P = 0.0094; MF vs. FM: P = 0.0022; FF vs. FM: P = 0.0326; FX vs. FM: P = 0.0011. All data are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 versus FM by two-way ANOVA with Sidak’s multiple comparisons test. Scale bars = 100 μm (b), 50 μm (a, e). Source data are provided as a Source Data file. The experiments in a, b, e, f were performed twice with similar results.