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Figure 10 | BMC Cell Biology

Figure 10

From: Regulation of endocytic trafficking of transferrin receptor by optineurin and its impairment by a glaucoma-associated mutant

Figure 10

The E50K mutant shows enhanced interaction with TfR. (A) E50K mutant shows enhanced interaction with TfR. HeLa cells were transfected with HA-tagged E50K mutant or wild type optineurin. After 30 hrs of transfection, lysates were prepared and immunoprecipitated with HA or control antibody and subjected to Western blotting with TfR, ubiquitin and HA antibodies. WCL, 5% whole cell lysate. (B) Interaction of E50K mutant optineurin with TfR is mediated through UBD. HeLa cells were transfected with HA-tagged E50K mutant or E50K-D474N mutant optineurin and immunoprecipitation was carried out as described in (A). WCL, 5% whole cell lysate. (C) E50K mutant colocalizes with ubiquitin. HeLa cells were transfected with GFP-tagged constructs of wild-type optineurin or its mutants and stained for endogenous ubiquitin. (D) E50K mutant colocalizes with ubiquitin in recycling endosomes. HeLa cells were co-transfected with GFP-tagged constructs of wild-type optineurin or E50K and HA-ubiquitin and stained with TfR and HA antibodies.

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