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. 2021 Apr 12;14:2475–2488. doi: 10.2147/OTT.S293180

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© 2021 Huang et al.

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XBP1 is a target of miR-516b-5p in ESCC cells. (A) The complementary sequence between miR-516b-5p and XBP1 was predicted by TargetScan software. (B) The transfection efficiency of miR-516b-5p mimics was assessed by RT-qPCR. (C) Dual-luciferase reporter assay was applied to test if miR-516b-5p bound to XBP1 in ESCC cells. (D and E) RIP assay was conducted to test the interaction between miR-516b-5p and XBP1 in ESCC cells. (F) RNA pull down assay was used to verify if miR-516b-5p interacted with XBP1 using biotinylated miR-516b-5p. Bio-NC was used as the control group. (G) The relative expression of miR-516b-5p was examined in 30 pairs of ESCC tissues and adjacent non-tumor tissues via RT-qPCR. RT-qPCR was used to measure the expression of miR-516b-5p in Eca109, KYSE150 and normal esophageal cell line HET-1A. *P<0.05, **P<0.01, ***P<0.001.