Fig. 5

CircNTRK2 acted as a sponge for miR-140-3p to abate it suppression of NRIP1 in ESCC cells. (a) Venn diagram showing the overlap of putative targets of miR-140-3p predicted by TargetScan, miRDB, DIANA-microT, DIANA-TarBase, and miRTarBase online databases. (b) The levels of NFYA, DAZAP2, CDK6, ACVR2B, VKORC1L1, and NRIP1 in Eca-109 and KYSE-150 cells transfected with miR-140-3p or miR-NC were detected by qRT-PCR. (c) The effect of miR-140-3p on NRIP1 protein expression was tested via western blot assay in Eca-109 and KYSE-150 cells. (d) The complementary sequences between NRIP1–3’UTR and miR-140-3p was shown. The red section represents their mutant (mut1 and mut2). (e) The luciferase activity of NRIP-wt, −mut1, −mut2, or -mut1 + 2 reporter was measured by dual-luciferase reporter assay in Eca-109 and KYSE-150 cells after transfection with miR-140-3p or miR-NC. (f) Expression difference of NRIP1 in ESCC tissues and normal tissues were predicted by GEPIA tool. (g) The level of NRIP in 56 pairs of ESCC tumor tissues and adjacent non-cancerous was examined via qRT-PCR. (h) The correlation between NRIP1 and miR-140-3p expressions in esophageal cancer was predicted in starbase Pan-Cancer Analysis Platform. (i) The protein level of NRIP1 was measured by using western blot assay in Eca-109 and KYSE-150 cells after transfection with circNTRK2 or circNTRK2 + miR-140-3p. (j and k) The correlation between NRIP1 and circNTRK2 or miR-140-3p was analyzed by Pearson test. *P < 0.05, **P < 0.01, ***P < 0.001