Figure 2
From: Dual function of partitioning-defective 3 in the regulation of YAP phosphorylation and activation
Par3 binds to the YAP PDZ-binding motif (PDZM) via its PDZ3 domain. (a) Par3 interacted with YAP. 293T cells were transiently transfected with the FLAG-Par3 and HA-YAP as indicated; co-immunoprecipitation (co-IP) and reverse-IP with anti-FLAG/HA antibodies were performed, and the cells were harvested for western blot analysis. (b) Endogenous Par3 bound to YAP in MDCK II cells. MDCK II cells were harvested and co-immunoprecipitated and reverse-immunoprecipitated with anti-Par3/YAP antibodies. (c) Modular structures of the three PDZ domain deletion mutations of Par3 and YAP. (d, e) Deletion of the Par3 PDZ3 eliminated its interaction with YAP. 293T cells were transfected with FLAG-Par3, FLAG-ΔPDZ1/2/3 mutants of Par3 and HA-YAP; co-IPs and reverse-IPs were conducted with anti-FLAG/HA antibodies. (f) Deletion of the PDZ motif of YAP also abrogated binding to Par3. 293T cells were transfected with FLAG-Par3, HA-YAP and HA-YAP/ΔPDZM; co-IPs were performed with anti-FLAG antibodies. (g) PDZ3 of Par3 was essential for the interaction, as determined by pull-down assay. 293T cells, which were transfected with HA-YAP, were lysed and incubated with glutathione-S-transferase (GST) and GST-tagged PDZ1/2/3 domains of Par3 bound to glutathione beads.
