Figure 4. Interaction with HSP90 is not required for clathrin binding by ACK2.

Myc-tagged ACK2 was transfected into COS7 cells for 48 h. The cells were then treated with geldanamycin (GA) or DMSO for either 10 or 30 min. The expressed ACK2 was immunoprecipitated (IP) with an anti-Myc antibody. The co-immunoprecipitated clathrin heavy chain and HSP90 were detected by immunoblotting with an anti-(clathrin heavy chain) antibody (top panel) and an anti-HSP90 antibody (third panel from the top) respectively. The immunoprecipitated ACK2 was detected by immunoblotting with an anti-ACK antibody (second panel from the top). The amounts of clathrin, ACK and HSP90 in the lysates are shown in the bottom three panels. The zero-time point sample represents cells without any treatment.