Fig. 5

FOXO3 was one direct target gene of miR-10b-3p. a-b FOXO3 was identified as potential regulatory targets of miR-10b-3p by considering the downregulation of genes using prediction tools and qRT-PCR. c The expression levels of FOXO3 mRNA and protein were measured by qRT-PCR and western blot analysis using GAPDH as the loading control after transfection of the miR-10b-3p mimic in the KYSE150 and KYSE450 cell lines, respectively. d The expression levels of FOXO3 mRNA and protein were measured by qRT-PCR and western blot analysis using GAPDH as the loading control after transfection of miR-10b-3p inhibitors in the KYSE30 and KYSE510 cell lines, respectively. e Dual-luciferase reporter assay. The relative luciferase activity was normalized to the Renilla luciferase activity assay after cotransfection with the miR-10b-3p mimic and miR-RB-REPORT constructs containing WT or MUT FOXO3 3’UTR region in KYSE150 and KYSE450 cell lines. Each experiment was performed in triplicate. Data are presented as the mean value ± SD. ^●, P < 0.05; ^●●, P < 0.01