Figure 2.

Enhancement of YAP/TAZ-Mediated Biological Actions by Parafibromin

(A) Wound healing assay was performed using AGS cells transiently transfected with an HA-TAZ^S89A vector or a control vector in the presence or absence of sgPF vector (left). The magnitudes of gap closures were quantified (middle). The levels of HA-TAZ^S89A and endogenous Parafibromin were determined by immunoblotting (right). Scale bars represent 100 μm.

(B) Colony formation assay of NIH3T3 cells transfected with the indicated plasmids.

(C) HEK293T cells were transfected with the indicated plasmids, and the cell proliferation was examined using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay 24 or 48 hr after transfection.

(D) qRT-PCR analysis of Cyr61 and Ctgf mRNA expression in Hrpt2^flox/flox;CAG-Cre-ER MEFs with or without 4-hydroxytestosterone treatment. Parafibromin expression was also determined by immunoblotting.

(E) Immunohistochemistry of the colon from Parafibromin conditional knockout (cKO) mice (Hrpt2^flox/flox;CAG-Cre-ER mice) with or without tamoxifen treatment. Yellow arrows indicate positive staining (Parafibromin-expressing colon epithelial cells, left panel; Cyr61-expressingcolon epithelial cells, right panel). H&E staining (upper, left); Parafibromin staining (lower, left); Ctgf staining (upper, right); Cyr61 staining (lower, right). Scale bars represent 20 μm.

Error bars, mean ± SD; n = 3; **p < 0.01 versus control vector (A–C), or 4-OHT(−) (D); ^‡p < 0.01 versus TAZ^S89A with control sgRNA (A–C). See also Figure S2.