The WW domains of Yap interact with the PPXY motif of Rx1.

<p>(A) Schematic illustration of the zebrafish Rx1 and Rx1 (ΔPPXY) constructs. A partial amino acid sequence including the PPXY motif of zebrafish Rx1 was aligned with the sequences of Rx from the indicated species. The PPXY motif (red lettering) is highly conserved among vertebrates. A detailed alignment of Rx family proteins can be found in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0097365#pone.0097365.s005" target="_blank">Fig. S5A</a>. (B) Co-immunoprecipitation analysis of HEK293T cells transiently expressing Myc-Rx1 that were co-transfected with empty vector (–), or vector expressing Yap (5SA), Yap (5SA/WW1^*, 2^*) or Yap (5SA/TEAD^*). Lysates were immunoprecipitated (IP) with anti-FLAG Ab to isolate Yap, followed by Western blotting (WB) with anti-Myc Ab to detect Myc-Rx1 (top), or with anti-FLAG Ab to detect FLAG-Yap (middle). (C) Co-immunoprecipitation analysis of HEK293T cells transiently expressing FLAG-Yap (5SA) that were co-transfected with empty vector (–), or vector expressing Rx1 or Rx1 (ΔPPXY). Lysates were IP’d using anti-FLAG Ab and subjected to WB with anti-Myc Ab to detect Rx1 (top), and with anti-FLAG Ab to detect Yap (middle). Bottom, WB analysis of total cell lysate using anti-Myc Ab to detect Rx1.