Fig. 3

CLDN4 induces EMT in vitro. a Phase-contrast micrographs of CLDN4 overexpressing cells, pEX2-NC cells and SGC-7901 cells. Scale bars = 50 μm. b, c The transcriptional and translational levels of EMT related markers. The real-time PCR and western blotting were performed at 48 h after the CLDN4 overexpressing cells treated with mimics of miR-596 or miR-3620-3p. d Immunofluorescence microscopy of the localization and expression of EMT and invasion markers in CLDN4 overexpressing cells. Scale bars = 50 μm. e The relative expression levels of CLDN4 in human GC tissues compared with their matched non-tumorous adjacent tissues. f Kaplan–Meier analysis of the correlation between CLDN4 expression levels and overall survival. g, h The relative expression levels of miR-596 and miR-3620-3p in human GC tissues compared with their matched non-tumorous adjacent tissues. i, j The correlation between CLDN4 transcriptional levels and miR-596 or miR-3620-3p transcriptional levels were measured in the same set of patients by Spearman correlation analysis. k Representative images of CLDN4, miR-596 and miR-3620-3p expression from GC tissues and non-tumorous adjacent tissues by ISH assays. Original magnification × 200. Scale bars = 100μm. Data are shown as mean ± s.d., n = 3. The data statistical significance is assessed by Student’s t-test and Wilcoxon’s Sign Rank Test is used to evaluate the differential expression between GC tissues and their matched non-tumorous adjacent tissues. *P < 0.05, **P < 0.01