Skip to main content
. 2016 Mar 16;7(17):24076–24087. doi: 10.18632/oncotarget.8119

Figure 4. MiR-1229 activates the Wnt/β-catenin signaling pathway by targeting GSK-3β, APC, and ICAT.

Figure 4

A. GSEA plot showing that miR-1229 expression was positively correlated with Wnt/β-catenin target gene signatures (REACTOME SIGNALING BY WNT and WILLERT_WNT_SIGNALING) in a published breast cancer dataset (TCGA). B. TOPflash or FOPflash luciferase reporter activity analyzed in the indicated cells. Cells transfected with TOPflash or FOPflash and Renilla pRL-TK plasmids were subjected to dual-luciferase assays 48 hours after transfection. The reporter activity detected was normalized by Renilla luciferase activity. C. The predicted miR-1229 target sequence in the 3′UTR of GSK3B, APC, and ICAT. The miR-1229 mutant (miR-1229-mut) contained three altered nucleotides in the seed sequence. D. Western blotting of GSK-3β, APC, and ICAT expression in MCF-7 and ZR-75-1 cells. α-Tubulin was used as the loading control. E. Luciferase assay of the indicated cells transfected with pGL-3UTR reporter and Vector, miR-1229, miRZip-Vector, miRZip-1229, or miR-1229-mut. F. RIP assay revealing the selective association between miR-1229 and GSK-3β, APC, or ICAT. Axin and GAPDH served as negative control. Bars represent the mean ± SD of three independent experiments. *P < 0.05.