Fig. 3: miR-1224-5p directly targets TNS4 in ESCC.

a The intersection of overexpressed genes in ESCC assayed by microarray and candidate targets predicted by the miRDB database. b TNS4 is overexpressed in ESCA by analysing the TCGA dataset using the GEPIA database. c, d Transcript levels of TNS4 in normal tissues and ESCC tissues from GSE53622 and GSE53624. e Western blotting was used to examine TNS4 protein expression in paired ESCC tissues and adjacent non-malignant tissues. f–i qRT-PCR and western blotting assays were used to measure the mRNA, and protein levels of TNS4. j The predicted binding site in the 3′-UTR of TNS4 targeted by miR-1224-5p. k, l Dual-luciferase activity assay showing the binding affinity between miR-1224-5p and the 3′-UTR of TNS4. m The transfection efficiencies of TNS4 siRNAs were detected by western blotting assay. n–q CCK-8 and colony formation assays were used to analyse the proliferation ability of transfected cells. r The negative correlation between miR-1224-5p and TNS4 was identified by analysing the TCGA dataset using the Starbase database. Data are represented as the mean ± SD, and experiments were performed in triplicate; * p < 0.05, ** p < 0.01, *** p < 0.001.