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. 2017 Feb 8;7:42319. doi: 10.1038/srep42319

Figure 3. CCNE2 is a direct downstream target of miR-664b-5p.

Figure 3

(A) The nucleotide sequences of miR-664b-5p and the complementary sequence in CCNE2 mRNA revealed a potential binding site. (B) Western blot analysis of cyclin E2 protein in miR-664b-5p overexpressing cells. GAPDH expression was used as the loading control. (C) qRT-PCR detection of CCNE2 mRNA expression in MDA-MB-436 and HCC1937 cells transfected with miR-664b-5p or miR-NC. (D) Relative luciferase activity was analysed after wild-type or mutant 3′-UTR reporter plasmids were cotransfected with miR-664b-5p or miR-NC in HCC1937 cells. (E) Fluorescence microscopy analysis of the expression of cyclin E2 by immunofluorescence. The green signal represents the staining of the cyclin E2 protein, and the blue signal represents the nuclear DNA staining by DAPI. *p < 0.05.