Skip to main content
. 2016 May 23;126(7):2495–2508. doi: 10.1172/JCI83361

Figure 6. miR-140-5p regulates key mediators of PAH pathology.

Figure 6

(A) Predicted PH-related direct targets of miR-140-5p. (B and C) qPCR (B) and protein expression by Western blot (C) showing expression of miR-140-5p targets ALK5, PDGFRA, and SMURF1 in PASMCs at 72 hours following transfection with miR-140-5p mimic (M), miR-140-5p inhibitor (I), or scramble control (S) (B: n = 6, C: n = 5, B and C: *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001, 1-way ANOVA with Tukey’s post-test correction, mean ± SEM). (D) Whole lung levels of SMURF1 protein correlate inversely with miR-140-5p (n = 38, P < 0.05, r = –0.36, Spearman rank). (E) Increased whole blood levels of SMURF1 mRNA were observed in patients with IPAH when compared with HV (IPAH n = 20, HV n = 16, *P < 0.05, unpaired 2-tailed Student’s t test, mean ± SEM). P, prevention; T, therapeutic. (F) Representative photomicrographs demonstrating immunoreactivity of SMURF1 in endothelial and SMCs of pulmonary vascular lesions in the transplanted lungs of patients with BMPR2 mutation–associated heritable PAH, associated PAH, and non-PAH control lung (photomicrographs representative of n = 26: APAH n = 8, IPAH n = 8, FPAH, n = 6, control n = 4). Original magnification, ×200 (hereditary PAH and control); ×400 (associated PAH). Scale bars: 50 μm. (G) Chromosomal location of putative miR-140-5p–binding site in the 3′ UTR of SMURF1. (H and I) Transfection of PASMCs with miR-140-5p inhibitor (H) and mimic (I) alter luciferase activity of SMURF1 3′ UTR construct at 24 hours (H and I: n = 4, **P < 0.01, ****P < 0.0001, 2-tailed Mann-Whitney U test, mean ± SEM).