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. 2019 Nov 18;130(1):94–107. doi: 10.1172/JCI128469

Figure 1. Post-transplant vascular injury is associated with endothelial repair gene expression.

Figure 1

(A) Hearts recovered 2 weeks after transplantation were immunostained for the EC marker CD31 or cadherin 5. Myocardial microvessel density was quantitated (CD31+ [PECAM], left; cadherin 5+, right). Apln+/y to male recipients (n = 12 biological replicates) experienced reperfusion injury alone; Apln+/y (n = 15) and Apln–/y (n = 14) to Apln+/+ female recipients experienced reperfusion and chronic alloimmune injury. HPF, high-power field. (B) Gaps in the arterial endothelium in cross section (left) and the fraction of cleaved caspase-3+ (aCasp) immunostaining among the CD31+ arterial endothelium (right) were quantitated among the samples from A. Endothelial repair gene expression among transplanted hearts was determined by qRT-PCR, and expressed relative to nontransplanted control hearts. (C) Gene expression among microdissected coronary arteries at 2 or 6 weeks after transplantation. Samples were pooled in pairs for analysis (at 2 weeks: Apln+/y to male recipients, n = 6 pairs; Apln+/y, n = 8, and Apln–/y, n = 7, to Apln+/+ female recipients; at 6 weeks: Apln+/y to male recipients, n = 5 pairs; Apln+/y, n = 5, and Apln–/y, n = 5, to Apln+/+ female recipients). (D) Gene expression among myocardium samples after transplantation (at 2 weeks: Apln+/y to male recipients, n = 12 biological replicates; Apln+/y, n = 15, and Apln–/y, n = 14, to Apln+/+ female recipients; at 6 weeks: Apln+/y to male recipients, n = 9 biological replicates; Apln+/y, n = 10, and Apln–/y, n = 10, to Apln+/+ female recipients). (E) Hearts recovered 2 weeks after transplantation were immunostained for endothelial CD31 (green) and ESM1 (red, arrows). Medium-sized to larger arterial cross sections are represented in the top panels, whereas myocardial microvessels are in the bottom panels. Scale bars: 50 μm. (F) ESM1 immunofluorescence quantitation among heart transplants in E; Apln+/y to male recipients (n = 12 biological replicates), Apln+/y (n = 15) and Apln–/y (n = 14) to Apln+/+ female recipients. Mean ± SEM; *P < 0.05, **P < 0.01, by 1-way ANOVA with Bonferroni’s post hoc test.