Fig. 4

MiR-146a inhibited ESCC cell invasion by targeting vimentin. a CE81^FN+ + CON stable cells were transiently transfected with 100 pmol/l of mimic-miR-146a or scramble microRNA (N.C.) using Lipofectamine™ for 48 h followed by measuring the expression of miR-146a by real-time PCR. b CE81^FN+CON cells harboring transiently transfected scramble (N.C.) or miR-146a used in (a) were further transiently transfected with pcDNA3.1-vimentin or vector, followed by measuring the level of vimentin using Western blotting. Total lysates in the same blotted PVDF membrane stained by Coomassie blue were used as the internal control. c The invasive ability of the cells used in (b) was evaluated by Transwell™ invasion assay. d CE81^FN+ + 146a stable cells were transiently transfected with anti-miR-146a or anti-scramble microRNA (anti-N.C.) followed by measuring the expression of miR-146a. e In parallel, the study was CE81^FN+miR-146a cells harboring anti-scramble (anti-N.C.) or anti-miR-146a were further transfected with si-Vimentin or si-RNA negative control (si–N.C.). The levels of vimentin and β-actin were investigated by Western blotting. Total lysates in the same blotted PVDF membrane stained by Coomassie blue were used as the internal control. f The invasive abilities of the cells used in (e) were measured by Transwell™ invasion assay. The numbers of the invaded cells on the bottom of the membrane were counted after incubating for 108 h. The quantitative data are shown. P values were obtained by Student’s t test, **P < 0.01, ***P < 0.001. The quantifications for (b) and (e) were derived from at least three independent biological repetitions (Additional file 1: Figure S7)