Fig. 4. PVT1 competitively binds miR-365 to regulate ELF4 expression. (A) Elevated miR-365 expression observed when PVT1 was knockdown in glioma cells U87 and U251, as detected by RT-qPCR. (B) ELF4 mRNA expression was downregulated when PVT1 was knockdown in glioma cells U87 and U251, as shown by RT-qPCR. (C) lncRNA PVT1 and ELF4 expression were positively correlated in glioma tissue samples, as analysed by Pearson’s correlation test. (D) Protein level of ELF4 was downregulated when PVT1 was knockdown in glioma cells U87 and U251, as shown by western blot. GAPDH was used for normalization. (E) PVT1 expression was downregulated in U87 cells after transfected with miR-365 mimics and upregulated after transfected with miR-365 inhibitor, as shown by RT-qPCR. (F) mRNA and protein level ELF4 expression were down-regulated in U87 cells after transfected with miR-365 mimics and and upregulated after transfected with miR-365 inhibitor, as demonstrated by western blot. GAPDH was used for normalization. (G) Bioinformatics analysis predicted that PVT1 specifically targeted miR-365, which was validated by dual-luciferase reporter assay. (H) ELF4 was a target for miR-365, as verified by bioinformatics analysis and dual-luciferase reporter assay. Data are presented as Mean±SD of three independent experiments. *p<0.05, **p<0.01.
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