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Figure 5


Fig. 5. T6BP and NDP52 colocalise and form a complex in vivo. (A) T6BP and NDP52 colocalise in vesicles around the Golgi complex in HeLa cells. Cells transiently overexpressing GFP-T6BP (a-d) or GFP-NDP52 (e-g) were double labelled with antibodies to NDP52 (a-d) and T6BP (e-h). The boxed areas in panels a and e are shown enlarged in adjacent panels b-d and f-h, respectively. Arrows indicate colocalisation. Bar, 10 µm. (B) Immuno-electron microscopy shows that endogenous T6BP and NDP52 colocalise on vesicles on the trans-side of the Golgi complex. Bar, 370 nm. (C) T6BP co-immunoprecipitates with NDP52. HeLa cells either untransfected (U) or overexpressing GFP-NDP52 (T) were lysed and immunoprecipitations were performed using no antibody (Blank) or anti-NDP52. Immunoprecipitates were immunoblotted using anti-T6BP antibodies. (D) T6BP and NDP52 interact in the mammalian two-hybrid assay. T6BP constructs were used as the bait and NDP52 constructs as the prey.





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