Figure 4.

USP36 interacts with cIAP1 and survivin in vivo. A, HEK-293 cells were transfected with Myc-cIAP1 and HA-USP36 individually or together. HA- or Myc-tagged proteins were immunoprecipitated from cell lysates and blotted with anti-HA and anti-Myc. B, HEK-293 cells were transfected with Myc-USP36 and HA-survivin individually or together. HA- or Myc-tagged proteins were immunoprecipitated from cell lysates and blotted with anti-HA and anti-Myc. C, HCT-8 cell lysates were immunoprecipitated with control IgG, anti-cIAP1, or anti-survivin antibody. Immunoprecipitates were analyzed via Western blot using indicated antibodies. D, HEK-293 cells were transfected with an empty vector or Myc-USP36, and protein interaction was assayed by immunoprecipitation with anti-cIAP1 or anti-survivin antibody and blotted with the indicated antibodies. E, HEK-293 cells were transiently transfected with Myc-USP36 alone or together with Flag-Bcl-xL, Flag-p65, or Flag-Mcl-1. Protein interaction was assayed by immunoprecipitation with anti-Flag magnetic beads and blotted with the indicated antibodies. F, HEK-293 cells were transiently transfected with Myc-USP36 alone or with Myc-USP36 and Flag-Mcl-1. Protein interaction was assayed using immunoprecipitation with anti-Myc beads and blotted with the indicated antibodies. G, HEK-293 cells were transiently transfected with Myc-XIAP alone or with Myc-XIAP and HA-USP36. Protein interaction was assayed using immunoprecipitation with anti-Myc magnetic beads and blotted with the indicated antibodies. HA, hemagglutinin; HEK, human embryonic kidney; IAP, inhibitor of apoptosis protein; Mcl-1, myeloid cell leukemia sequence 1; USP36, ubiquitin-specific protease 36.