Fig. 4

SMYD4 suppressed Wnt signaling by inhibiting the nuclear localization of βcatenin. (A) RNAseq differential gene volcano map between SMYD4 overexpressed and control group. The GO analysis was performed among up-regulated genes (B)and down-regulated genes(C). (D)TOP5 up- and down-regulated gene sets after GSEA analysis and the enrichment results of WNT signalling pathway(E). (F)The βcatenin expression level in the nucleus and cytoplasm detected by nucleocytoplasmic separation western blot. (G)The TOP/FOP luciferase reporter assay in SMYD4overexpressed CAL-51 cells and in SMYD4 knock-down MDAMB-231 cells. (H) The localization transformation of βcatenin visualized by IF. (I)The expression levels of downstream target genes of Wnt/βcatenin signaling, including MYH9,N-cadherin, Vimentin, C-myc, Cyclin D1 and Survivin detected by western blot. All experiments were repeat three times (n = 3), compared variances using F test and analyzed using an unpaired t-test, error bars represent SD. **p < 0.01;***p < 0.001; ****p < 0.0001