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. 2013 Apr 24;123(5):2143–2154. doi: 10.1172/JCI64365

Figure 2. NRAS is a target gene of miR-146a in HUVECs.

Figure 2

(A) NRAS mRNA (qRT-PCR) and (B) protein levels (Western blot) in HUVECs transfected with pre-miR-146a and pre-miR-control. (C) Luciferase activity from NRAS 3′UTR WT reporter plasmid and mutated NRAS 3′UTR cotransfected into HEK293T cells with pre-miR-146a or pre-miR-control 48 hours after transfection. (D) BrdU incorporation, (E) FACS analysis for apoptosis by annexin V–PI staining, and (F) DNA fragmentation analysis in HUVECs transfected with NRAS or control siRNA for 48 hours. (G) Representative images of laser-induced choroidal neovascularization 7 days after transfection with NRAS or control siRNA injected intravitreously (n ≥ 8 eyes/condition; 4 lesions/eyes). Dashed outlines denote lesion area. Scale bars: 100 μm. (H) Quantification (by ImageJ) of the green signal present in lesion area. CNV, choroidal neovascularization. (I) NRAS mRNA level in HUVECs stimulated with 16K PRL (50 nM, 8 hours), with pretransfection (48 hours) with anti-miR-control or anti-miR-146a. All data are mean ± SD (n ≥ 3) or mean ± SEM (H). *P < 0.05 vs. respective control. See also Supplemental Figure 2.