Fig. 3: ZIC5 increases Wnt/β-catenin signaling by potentiating the interaction between β-catenin and TCF4.
From: AR-regulated ZIC5 contributes to the aggressiveness of prostate cancer
A, B C4-2B (A) and 22RV1 (B) cells were transfected with sh-ZIC5 or oe-ZIC5. Relative expression of Wnt/β-catenin target genes (c-Myc, MMP2, and MMP7) was measured by RT-qPCR. *P < 0.05, relative to sh-NC or oe-NC. C, D C4-2B (C) and 22RV1 (D) cells transfected with sh-NC or sh-ZIC5 and treated with LiCl (20 mmol/L) for 24 h. Relative expression levels of c-Myc, MMP2 and MMP7 were analyzed by RT-qPCR. *P < 0.05, vs. sh-NC, #P < 0.05 vs. sh-ZIC5. E TCF/LEF Luciferase reporter assay results depicting Wnt/β-catenin signaling activity in C4-2B and 22RV1 cells after ZIC5 knockdown or overexpression. *P < 0.05 vs. sh-NC or oe-NC. F Western blot analysis of overall β-catenin expression in C4-2B and 22RV1 cells transfected with sh-NC or sh-ZIC5. G Western blot assessment of β-catenin protein levels in the cytoplasm and nucleus of C4-2B and 22RV1 cells treated as indicated. H The cellular distribution of β-catenin assessed through immunofluorescence staining. Scale bars, 20 μm. I Co-IP analysis of interactions between ZIC5 and β-catenin/TCF4 in C4-2B and 22RV1 cells transfected with Flag-tagged ZIC5 or control vector. J, K Flag-tagged ZIC5 or control vectors were transfected into 293T cells and Co-IP was performed on nuclear extracts probed with anti-TCF4 (J) or anti-β-catenin (K) antibodies.
