Fig. 2

Predication and validation of vimentin as a target of MiR-146a gene in ESCC cells. a The expression level of miR-146a in CE81^FN+ + CON and CE81^FN+ + 146a stable cells was evaluated by real-time PCR. The data was analyzed by ΔCT method and the relative expression was normalized to the internal control U54 gene expression. b The expression levels of the six overlapped target genes of miR-146a predicted by three software programs, i.e., Targetscan, Microcosm and miRNAMap were measured in CE81^FN+CON and CE81^FN+146a stable cell lines by real-time PCR. c The mRNA expression levels of vimentin in CE81^FN+ and CE81^FN− cells were determined by RT-PCR. d The protein levels of vimentin in CE81^FN+ and CE81^FN− cells were determined by Western blotting. β-actin was used as the internal control. e Vimentin was labelled by FITC conjugated anti-vimentin antibody in CE81^FN+ and CE81^FN− cells by IFA assay. The cells with fluorescent vimentin expression were shown and quantified by the fluorescent microscopy (×400). Scale bar: 100 μm. P values were obtained from the Student’s t test. Statistically significant difference was indicated (*P < 0.05, ***P < 0.001)