Research Paper Volume 13, Issue 8 pp 12239—12257

Figure 8. Depletion of miR-503-5p inhibits the formation of atherosclerotic plaques in vivo. (A) Expression of miR-503-5p (normalized to U6), and TGF-β1, smad7, smurf1, and smurf2 (normalized to GAPDH) in plasma-EVs, macrophage-EVs, and arterial tissues of ApoE-/- mice (n = 3) and WT mice (n = 3) determined by RT-qPCR. (B) Protein levels of TGF-β1, smad7, smurf1, and smurf2 (normalized to GAPDH) in plasma-EVs, macrophage-EVs, and arterial tissues of ApoE-/- mice (n = 3) and WT mice (n = 3) determined by Western blot analysis. (C) Lipid deposition in mouse arterial tissues detected by Oil red O staining; Scale bar = 50 μm. (D) EVs derived from macrophages with or without ox-LDL treatment were separately injected into the tail vein of C57BL6 WT mice at dose of 100 μg for each. One week later, EVsinjected mice were injected with lentivirus expressing anti-miR-503-5p and anti-miR-503-5p NC. (E) The lipid plaque of arterial tissues of C57BL/6J WT mice (n = 12) detected by Oil red O staining. (F) Expression of smad7, smurf1, and smurf2 (normalized to GAPDH) in arterial tissues of C57BL/6J WT mice (n = 12) determined by Western blot analysis. Values obtained from three independent experiments in triplicate were analyzed by unpaired t test between two groups and by one-way ANOVA followed by Tukey's post hoc test among three or more groups. * p < 0.05 compared with WT mice or mice injected with EVs derived from macrophages without ox-LDL treatment; # p < 0.05 compared with mice injected with EVs derived from macrophages with ox-LDL treatment followed by lentivirus expressing anti-miR-503-5p NC.