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Description
Ema localizes to endosomes. Third instar larval garland cells expressing Ema-GFP fusion protein and stained for the late endosomal and lysosomal protein Spinster (red) and plasma membrane and nuclei (blue). Ema co-localizes with some late endosomes. Wandering third instar larvae were dissected in ice-cold PBS, equilibrated in HL-3 buffer at room temperature, incubated in 0.7% HRP for 5 min at room temperature, rinsed thoroughly and fixed with Bouin's solution for 5 min. Blocking and antibody incubation were performed in PBS containing 0.1% Triton X-100. Anti-Spinster antibody (Sweeney and Davis, 2002) used at 1:250. NeuroTrace 640/660 deep red fluorescent Nissl (Invitrogen) used at 1:500 was used to stain nuclei. Cy5-conjugated anti-HRP (Jackson Immunoresearch Laboratories) at 1:1000 was used to stain plasma membrane. Cy3-conjugated secondary antibody (Jackson Immunoresearch Laboratories, Inc.) at 1:1,000. 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 3B 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:13477, Drosophila melanogaster, garland cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J09K48ZG
Type
image
Identifier
ark:/20775/bb3351347z
Language
No linguistic content
Subject
Endocytosis Late endosome Endosome Plasma membrane Nucleus Garland cell Drosophila melanogaster Cell Image Library Group ID: 6022
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