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. 2018 Feb 15;1:1–15. doi: 10.1016/j.isci.2018.01.003

Figure 7.

Figure 7

Synergism between Nuclear TAZ and the Wnt Signaling Pathway by Parafibromin

(A) HEK293T cells were transfected with a TOP-tk luciferase reporter together with indicated plasmids. TCLs were subjected to luciferase assay and immunoblotting analysis with indicated antibodies.

(B) HEK293T cells or AGS cells were cultured in a low-density (L) or a high-density (H) condition, and TOP-tk luciferase reporter activity was measured.

(C) Total cell lysates from (B) were subjected to immunoblotting.

(D) HEK293T cells were transfected with indicated plasmids. TCLs and the anti-FLAG immunoprecipitates were subjected to immunoblotting with the indicated antibodies.

(E) HEK293T cells were transfected with a TOP-tk luciferase reporter together with indicated plasmids. Cells were then treated with Wnt3a-conditioned medium or control medium. TCLs were subjected to luciferase assay and immunoblotting with the indicated antibodies.

(F) AGS cells were transfected with a TOP-tk luciferase reporter together with indicated plasmids. TCLs were subjected to luciferase assay and immunoblotting with the indicated antibodies.

(G) AGS cells were transfected with indicated vectors. Luciferase assay (left) and qRT-PCR (right) experiments were performed.

(H) AGS cells were transfected with indicated plasmids. Anti-FLAG immunoprecipitates from TCLs were subjected to immunoblotting with the indicated antibodies.

(I) An HA-β-cateninS33Y vector was transfected together with a Myc-TAZ or Myc-TAZΔTEAD vector into HEK293T cells, in which expression of endogenous Parafibromin was acutely inhibited by using CRISPR/CAS9-mediated gene knockout technique. Anti-HA (left) or anti-Myc (right) immunoprecipitates were prepared from TCLs and subjected to immunoblotting with the indicated antibodies. β-catS33Y; β-cateninS33Y.

Error bars, mean ± SD; n = 3; *p < 0.05 versus control vector (A and E); **p < 0.01 versus control vector (A, E–G), low cell density (B), or control vector with TAZ (G); ‡p < 0.01 versus PF with control vector (G, left), PF (G, right), or PF with TAZ (G).

See also Figure S7.