Figure 5.

Differential Modes of Parafibromin Binding between YAP and TAZ

(A) HEK293T cells were co-transfected with indicated plasmids. Anti-FLAG IP from cell lysates and TCL were analyzed by immunoblotting with anti-FLAG and anti-HA antibodies (left). A schematic of Parafibromin and its deletion mutants (right). The three tyrosine dephosphorylation sites (Y290/293/315) are indicated.

(B) HEK293T cells were transfected with an FLAG-PF or FLAG-phosphoresistant (PR)-PF vector together an HA-TAZ vector. Anti-FLAG IP from cell lysates and TCL were subjected to immunoblotting with anti-FLAG and anti-HA antibodies.

(C) HEK293T cells were transfected with a TEAD luciferase reporter together with indicated plasmids. Total cell lysates were subjected to luciferase assay and immunoblotting analysis with indicated antibodies.

(D) Wound healing assay of AGS cells co-transfected with a TAZ^S89A vector and a PF or PR-PF vector. Scale bars represent 100 μm.

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

(F) HEK293T cells were transfected with a TEAD luciferase reporter together with indicated plasmids. Total cell lysates were subjected to luciferase assay and immunoblotting with indicated antibodies.

Error bars, mean ± SD; n = 3; **p < 0.01 versus control vector with TAZ (C), PF with TAZ^S89A (D), control vector with YAP2δ (F), or control vector with YAP1δ (F); ^‡p < 0.01 versus PF with TAZ (C), PF with YAP2δ (F), or PF with YAP1δ (F).

See also Figure S5.