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. 2015 Aug 21;94(33):e1327. doi: 10.1097/MD.0000000000001327

FIGURE 5.

FIGURE 5

MicroRNA (MiR)-664 suppresses PLP2 expression via direct targeting of the 3′UTR. (A) Predicted miR-664 target sequence in the 3′UTR of PLP2 (PLP2-3′UTR) and PLP2-3′UTR mutant containing 4 altered nucleotides in the seed sequence (PLP2-3′UTR-mut). (B) Western blot analysis of PLP2, p-AKT, AKT, p21, cyclin D1, and PCNA expression by miR-664 overexpression or miR-664 inhibition. β-actin served as the loading control. (C) Western blot analysis of GFP expression in indicated cells. (D) Luciferase assay of indicated cells transfected with the pGL3-PLP2-3′UTR reporter with increasing amounts (10, 50 nM) of miR-664 mimic or miR-664 inhibitor. (E) Luciferase assay of indicated cells transfected with pGL3-PLP2-3′UTR or pGL3-PLP2-3′UTR-mut reporter with increasing amounts (10, 50 nM) of miR-664 oligonucleotides. (F) Analysis of expression (up) and correlation (down) of miR-664 and PLP2 expression in 8 freshly collected human CMM tissues. Each bar represents the mean ± standard deviation of 3 independent experiments. ∗P < 0.05. 3′UTR = untranslated region, AKT = protein kinase B, CMM = cutaneous malignant melanoma, p-AKT = AKT phosphorylation, PLP2 = proteolipid protein 2.