C. elegans TGN-38-GFP localization depends upon retromer, RME-1, CDC-42, TOCA-1/TOCA-2, PAR-6, PKC-3, and WVE-1/WAVE. (A) An alignment of intracellular domains of rat TGN38 and C. elegans TGN-38 is shown. Conserved endocytosis signal Y314 is marked with an arrowhead. (B–G and N–R) Confocal images of C. elegans intestinal cells expressing GFP-tagged TGN-38 in the marked genotypes. (H–M and S–W) Endocytosis-defective Y314A mutant version of TGN-38-GFP in the indicated genetic backgrounds. Note the increase in TGN-38-GFP intensity upon loss of retromer, RME-1, CDC-42, TOCA-1/TOCA-2, PAR-6, PKC-3, or WVE-1. The endocytosis-defective Y314A version of TGN-38-GFP was unaffected, indicating the requirement for these trafficking factors in postendocytic trafficking of TGN-38. (Scale bars, 10 µm.)