Under copyright Constraint(s) on Use: This work is protected by the U.S. Copyright Law (Title 17, U.S.C.). Use of this work beyond that allowed by "fair use" or any license applied to this work requires written permission of the copyright holder(s). Responsibility for obtaining permissions and any use and distribution of this work rests exclusively with the user and not the UC San Diego Library. Inquiries can be made to the UC San Diego Library program having custody of the work. Use: This work is available from the UC San Diego Library. This digital copy of the work is intended to support research, teaching, and private study.
Rights Holder and Contact
UC Regents
Description
Ema localizes to endosomes. Third instar larval garland cells expressing Ema-GFP fusion protein and stained for the early endosomal protein Rab5 (red) and plasma membrane and nuclei (blue). Ema does not co-localize with early 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 in 4% paraformaldehyde in PBS for 30 min. Blocking and antibody incubation were performed in PBS containing 0.1% Triton X-100. Anti-Rab5 (Wucherpfennig et al., 2003) at 1:50. 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 3C 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:13478, Drosophila melanogaster, garland cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0BP01G4
Type
image
Identifier
ark:/20775/bb9426487g
Language
No linguistic content
Subject
Endocytosis Plasma membrane Nucleus Early endosome Garland cell Endosome Drosophila melanogaster Cell Image Library Group ID: 6022
If you're wondering about permissions and what you can do with this item, a good starting point is the "rights information" on this page. See our terms of use for more tips.
Share your story
Has Calisphere helped you advance your research, complete a project, or find something meaningful? We'd love to hear about it; please send us a message.
