Figure 1: miRNA and Target Profiling in Radiated Versus Nonradiated Human Arteries

(A) Expression of 11 vascular disease-related microRNAs (miRNAs) in radiated (R) versus nonradiated (NR) arteries. n = 12 per group. Mean ± SEM. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. (B) miR-29b was down-regulated and miR-146b upregulated in radiated versus nonradiated arteries harvested at microvascular tissue transfers. n = 15. ∗p < 0.05; ∗∗∗p < 0.001. (C) Fluorescent in situ hybridization of miR-29b showing its expression in the tunica media of nonradiated vessels. Pentraxin-3 (PTX3) and dipeptidyl-peptidase 4 (DPP4) protein expression is increased throughout the vessel wall after radiation. ACTA2 = smooth muscle actin; DAPI = 4′,6-diamidino-2-phenylindole. Bars, 100 μm. (D) Quantification of staining for miR-29b target protein markers. n = 3 in each group. HPF = high-power field. **p < 0.01 in 1-way analysis of variance. (E) miR-29b target PTX3 was up-regulated after irradiation. n = 10. ∗∗p < 0.01. (F) miR-29b target DPP4 was upregulated after irradiation. n = 10. *p < 0.05, **p < 0.01, ***p < 0.001 in paired-sample t-tests. (G) miR-29b mimics significantly inhibited DPP4 and PTX3 luciferase activity compared with scrambled control (scr) oligonucleotides. Mean ± SEM. **p < 0.01, ***p < 0.001 in Student’s t-test of scrambled versus miR-29b mimic. EV = empty vector; mut = mutated seed sequence; Neg ctrl = negative control.

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