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. 2019 Dec 23;19(2):1409–1417. doi: 10.3892/ol.2019.11241

Figure 4.

Figure 4.

MiR-3677 suppresses TLE3 expression by directly targeting the TLE3 3′-UTR and altered levels of associated proteins in MCF-7 and ZR-75-1 cells. (A) Predicted miR-3677 target sequence in the 3′-UTR of TLE3 and positions of three mutated nucleotides (red) in the 3′-UTR of miR-3677 (miR-3677 mut). (B) TLE3 mRNA expression in MCF-7 and ZR-75-1 cells transfected with miR-3677 or miR-3677-in or miR-3677-mut were detected using RT-qPCR analysis. (C) Luciferase reporter assay of MCF-7 and ZR-75-1 cells transfected with the pGL3-TLE3-3′-UTR reporter and miR-3677 or miR-3677-in or miR-3677-mut or NC. (D) TLE3 protein expression in MCF-7 and ZR-75-1 cells transfected with miR-3677 or miR-3677-in or miR-3677-mut were detected by western blotting analysis. α-tubulin served as the loading control. (E) RT-qPCR analysis of expression of cyclin D1 and c-myc in MCF-7 and ZR-75-1 cells. (F) Cyclin D1 and c-Myc were measured by western blotting in MCF-7 and ZR-75-1 cells. α-tubulin served as the loading control. *P<0.05 miR-3677 vs. vector or miR-3677-in vs. NC. miR, microRNA; TLE3, transducin-like enhancer of Split3; UTR, untranslated region; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; NC, negative control; in, inhibitor.