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Ema localizes to endosomes. Third instar larval garland cells expressing Ema-GFP fusion protein with endosomes labeled with a 5-min pulse of the fluid-phase endocytic tracer avidin-Cy3 followed by an additional 5-min chase. Ema localizes to functional endosomes. Third instar larvae were dissected in ice-cold PBS, equilibrated in the Schneider medium for 5 min at room temperature, pulsed with 10 µg/ml of Extravidin-Cy3 (Sigma-Aldrich) at room temperature for 5 min, rinsed thoroughly, and chased in Schneider medium. Samples were fixed in 4% PFA in PBS for 10 min at room temperature. Confocal images were acquired with a confocal microscope (model C1; Nikon) and accompanying EZ-C1 software using argon (excitation at 488 nm) and HeNe (excitation at 543 and 633 nm) lasers and a 60x Plan-Apochromat NA 1.4 objective (Nikon) at room temperature. Samples for each experiment were processed using the same confocal gain setting. Image corresponds to Fig 3A in Kim et al. J Cell Biol. 188: 717-734. 2010. Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp) Kim, Sungsu; Wairkar, Yogesh P.; Daniels, Richard W.; DiAntonio, Aaron (2021). CIL:13476, Drosophila melanogaster, garland cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0639NFF
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