Fig. 4

MINDY1 deubiquitylates YAP. A T24 cells transfected with the indicated siRNA were treated with MG132 for 6 h before collection. YAP was immunoprecipitated with anti-YAP and immunoblotted with anti-HA. B Immunoblotting to detect the ubiquitination of YAP in HEK293 cells co-transfected with Flag-YAP, HA-Ubiquitin and Myc-MINDY1 (wild type or C137A). C MINDY1 removed the ubiquitin chain of YAP in a dose-dependent manner. D, E K48 or K63 Ub was co-transfected with Flag-YAP and Myc- MINDY1 into HEK293 cells. After treatment with 10 μM MG132 for 6 h, cell lysates were subjected to ubiquitination assay and the ubiquitination level of YAP was detected by HA antibody