Figure 12
The induction of cell death in RGC-5 cells by E50K mutant requires the UBD. (A) RGC-5 cells were transfected with wild type optineurin or E50K mutant or their UBD mutants and percentage of cells undergoing cell death were quantified in transfected and untransfected cells. Data represent mean ± SD of 3 independent experiments. (B) E50K mutant colocalizes with TfR in RGC5 cells. GFP-tagged optineurin or its mutants were transfected in RGC-5 cells and stained for TfR. E50K mutant optineurin forms large vesicular structures that co-localize strongly with TfR indicated by yellow colour in the merged image. (C). E50K mutant causes reduced uptake of transferrin in RGC-5 cells. RGC-5 cells were transfected with GFP- tagged wildtype optineurin or its mutants and incubated with Alexa-546 conjugated transferrin for 20 min. Cells were then fixed and observed with a confocal microscope. The uptake of labelled transferrin was quantitated by measuring the fluorescence intensity in expressing (filled bar) and non-expressing (empty bar) cells. Bar; 10 μm.