FIG 5.

HBZ abrogates the interaction of CUL1 with Skp1 but not with Rbx1. HEK293T cells were cotransfected with 2 μg of a plasmid expressing Myc-Rbx1 (A) or Myc-Skp1 (B) with (+) or without (−) 1 μg of a plasmid expressing HA-CUL1 and 5 μg of a plasmid expressing HBZ-FLAG, as indicated. After 36 h, cell extracts were prepared and subjected to immunoprecipitation using anti-HA antibodies, followed by immunoblot analysis with anti-Myc and anti-HA antibodies. Total protein levels in whole-cell lysates were analyzed by immunoblotting using anti-Myc, anti-HA, anti-FLAG, and anti-α-tubulin antibodies. The intensity of each band was quantified and graphed (*, P < 0.05). WCL, whole-cell lysates; IB, immunoblotting; IP, immunoprecipitation.