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. 2018 Jun 5;18(2):1616–1622. doi: 10.3892/mmr.2018.9131

Figure 4.

Figure 4.

TCF4 was a direct target of miR-4695-5p in OS cells. (A) Predicted binding site of miR-4695-5p in the 3′-UTR of TCF4 mRNA. (B) Luciferase reporter assays indicated that overexpression of miR-4695-5p suppressed the luciferase activity in Saos-2 cells transfected with WT 3′-UTR. (C) RT-qPCR analysis indicated that overexpression of miR-4695-5p inhibited the mRNA levels of TCF4 in Saos-2 and U2OS cells. (D) RT-qPCR analysis showed that the expression of miR-4695-5p was reversely correlated with that of TCF4 in OS tissues. (E) Overexpression of miR-4695-5p suppressed the protein level of TCF4 and activation of Wnt/β-catenin pathway in Saos-2 and U2OS cells. The protein level of TCF4, Cyclin D1 and MYC were measured by western blot. GAPDH was chose as a loading control. The results represented three independent experiments and were expressed as mean ± SD. *P<0.05 and **P<0.01 vs. the control group. TCF4, transcription factor 4; OS, osteosarcoma; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.