Fig. 6

Mechanism of tumor suppressive function of CLDN7 in ccRCC. (A) Heatmap representation of differentially expressed genes identified by RNA-Seq between Caki-1 CLDN7 cells (n = 3) and Caki-1 Control cells (n = 3). (B) Statistics of KEGG pathway enrichment. The y-axis corresponds to KEGG Pathway, and the x-axis shows the GeneRatio. The color of the dot represent adjusted p value (padj), and the size of the dot represents the number of differentially expressed genes mapped to the reference pathways. (C) Validation of differentially expressed genes by qRT-PCR. Comparison of mRNA expression of genes in pathways of cancer (BCL2, HIF1A, GLI-1, ITGB-1, p21 and AR) and genes in EMT-related pathway (TGFB1, E-cadherin, N-cadherin, Vimentin and TWIST1) between Caki-1 CLDN7 cells and Caki-1 Control cells. All data are shown as means ± SD. (D) a. Western blot assay (left) and statistical analysis (right) of CLDN7, BCL2, cleaved-PARP1 and cleaved-Caspase 3 expression in Caki-1 and A498 cells, while overexpression of CLDN7 compared with control group. b. IHC assay of BCL2, cleaved-Caspase 3, Ki-67 and CLDN7 expression in xenografts formed by Caki-1 CLDN7 and Control cells. Scale bar, 100 μm. (E) a. Western blot assay and statistical analysis of E-cadherin, N-cadherin and Vimentin expression in CLDN7 overexpressed Caki-1 and A498 cells, comparing with control cells. b. IHC assay of E-cadherin, N-cadherin, TGFB1 and CLDN7 in xenografts formed by Caki-1 CLDN7 and Control cells. Scale bar, 100 μm. N.S, not significant. *p < 0.05, **p < 0.01, Student’s Test