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. 2014 Feb 19;5(10):2999–3011. doi: 10.18632/oncotarget.1626

Figure 3. β-Catenin nuclear accumulation and transcriptional transactivation by PI3K are required for β-Catenin mediated transcription.

Figure 3

(A) TOP-flash luciferase reporter assay of 293T cells expressing a stabilized mutant β-Catenin protein (Δ45-β-CateninER-myc) whose nuclear accumulation is inducible by 4-OH-tamoxifen (4-OHT) treatment. Cells treated for 12 hours with the 4-OHT concentrations indicated or vehicle (EtOH). (B) TOP-flash luciferase reporter assay of CHO-1 and HeLa cells expressing Δ45-β-CateninER-myc treated for 12 hours with 500 nM 4-OHT (C) Immunofluorescence microscopy of 293T and CHO-1 cells expressing Δ45-β-CateninER-myc treated with 4-OHT or vehicle for 12 hours. DAPI marks nuclei. (D) TOP-flash luciferase reporter assay of 293T, CHO-1 and HeLa cells expressing Δ45-β-CateninER-myc treated with 500 nM 4-OHT or vehicle and 25 μM LY or vehicle. (E) Immunofluorescence microscopy of cells transfected with Δ45-β-CateninER-myc and pTOP-GFP, treated with 25 μM LY and 500nM 4-OHT or vehicle for 12 hours. DAPI marks nuclei. (F) Western blot analysis of 25 μg whole cell protein lysates of 293T and CHO-1 cells transfected with Δ45-β-CateninER-myc and pTOP-GFP as in figure (E) and treated with the indicated substances for 12 hours.