Fig. 1

CLDN4 is identified as a target of miR-596 and miR-3620-3p. a Hierarchical clustering analysis of mRNAs, lncRNAs, and miRNAs that were differentially expressed between GC tissues and non-tumorous adjacent tissues (>2.0-fold; P < 0.05; filtered to show the top 30 up-regulated or down-regulated results for mRNAs and lncRNAs). Expression values are represented in shades of red and green, indicating expression above and below the median expression value across all tissues, respectively. b The mRNA-lncRNA -miRNA networks in the GC. The networks include cell adhesion pathway, pathway in cancer, tight junction pathway, and cell cycle pathway. Genes colored in green are protein-coding RNAs associated with GC. Genes colored in blue are lncRNAs and genes colored in red are miRNAs associated with GC. c Predicted binding sites for miR-596, miR-3620-3p, and miR-4292 on the CLDN4 transcript. The white nucleotides are the seed sequences of miRNAs. d Real-time PCR analysis of CLDN4 expression in GC cells treated with mimics of miR-596, miR-3620-3p, miR-4292, and negative control. e GC cell line SGC-7901 was transfected with the mimics of miR-596, miR-3620-3p, miR-4292, and negative control. Reduced CLDN4 expression was shown by western blotting analysis and normalized to β-tubulin. f Luciferase activities were measured in GC cells co-transfected with luciferase reporter containing CLDN4 and the mimics of miR-596, miR-3620-3p, miR-4292, or mutant. Data are presented as the relative ratio of renilla luciferase activity and firefly luciferase activity. g The relative expressions of CLDN4 were determined by real-time PCR and western blotting. Data are shown as mean ± s.d., n = 3. The data statistical significance is assessed by Student’s t-test. *P < 0.05, **P < 0.01