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Figure 1 | BMC Cell Biology

Figure 1

From: UIM domain-dependent recruitment of the endocytic adaptor protein Eps15 to ubiquitin-enriched endosomes

Figure 1

FLAG-Eps15 is recruited to endosomes enriched in ubiquitinated ErbB2 in a UIM-domain-dependent manner. A, B. FLAG-Eps15-transfected SK-BR-3 cells were treated with DMSO (A) or 5 μM GA (B) for 4 hours, and then processed for IF, detecting FLAG-Eps15 with anti-FLAG antibodies and endogenous ErbB2 with anti-ErbB2 antibodies. C-F. COS-7 cells transfected with FLAG-Eps15 or FLAG-Eps15-ΔUIM, together with GFP or ErbB2-GFP as indicated, were treated with 5 μM GA for 4 hours and then processed for IF, detecting FLAG-Eps15 constructs with anti-FLAG antibodies. G. Colocalization of FLAG-Eps15 or FLAG-Eps15-ΔUIM with EEA1 in COS-7 cells co-expressing either GFP or GFP-ErbB2 and treated with DMSO or 5 μM GA for 4 hours was quantified using NIH ImageJ. The Manders’ overlap coefficient is shown. Values shown are the average of at least 10 cells in each of 3 experiments, +/- standard error of the mean (SEM). The data were compared using a one-way ANOVA test (***p < 0.0001). Scale bars; 10 μm.

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