SNX16 localizes to Rab5-positive early endosomes. (A) Kc167 cells transfected with Snx16-GFP were treated with wortmannin and then fixed and imaged by confocal microscopy. Single confocal sections are shown. (B) S2 cells transfected with the indicated constructs were spread on concanavalin A, fixed, and imaged. 2D projections of confocal stacks are shown. (C) Kc167 cells transfected with human transferrin receptor and the indicated constructs were incubated with Alexa Fluor 546–labeled transferrin for the indicated times and then fixed and imaged. Single confocal sections are shown. (D) Nwk localizes to a subset of SNX16 puncta. S2 cells were transfected with Snx16-mRFP and nwk-GFP and spread on concanavalin A before fixation and imaging. A 2D projection of a confocal stack is shown. Arrows indicate overlapping puncta. Bars, 10 µm.