Figure 9
Optineurin interacts with TfR through UBD. (A) Endogenous optineurin interacts with endogenous TfR. Endogenous optineurin was immunoprecipitated from HeLa cells using anti-optineurin or control antibody. Immunoprecipitates were resolved in SDS-PAGE and subjected to Western blotting with TfR and optineurin antibodies. (B) UBD of optineurin is essential for interaction with TfR. HeLa cells were transfected with HA-optineurin or HA-D474N mutant optineurin. After 30 hrs of transfection, lysates were immunoprecipitated with anti-HA or control antibody and subjected to Western blotting with TfR, ubiquitin and HA antibodies. (C) TfR was immunoprecipitated from optineurin-overexpressing and control cells. Western blot of the immunoprecipitate was done with ubiquitin, TfR and optineurin antibodies. WCL, 5% whole cell lysate.