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Description

The endosomal compartment is enlarged in the ema[1] mutant. Colocalization between DVGLUT (red) aggregates and the endosomal ESCRT protein Hrs (green, also known as Vps27) in motoneuron cell bodies of the ventral nerve cord of the ema[1] mutant. Wandering third instar larvae were dissected in ice-cold PBS and fixed with Bouin's solution for 5 min. Blocking and antibody incubation were performed in PBS containing 0.1% Triton X-100. anti-DVGLUT (Daniels et al., 2004) antibody was used at 1:10,000; anti-HRS-FL (Lloyd et al., 2002) was used at 1:1000. Secondary antibodies (Alexa 488-/Cy3-conjugated) were used at 1:1000. After staining, specimens were equilibrated in 70% glycerol in PBS and mounted with VectaShield (Vector Laboratories). 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 Figure 2A, bottom 3 panels in Kim et al. J Cell Biol. 188: 717-734. 2010.

Biological Sources

NCBI Organism Classification
Drosophila melanogaster
(Common fruit fly)
Cell Type
motor neuron
Cell Line
ema[1]
Cellular Component
endosome

Biological Context

Biological Process
receptor-mediated endocytosis
endosome to lysosome transport
endosome transport
Molecular Function
zinc ion binding

Attribution

Names
Sungsu Kim
Yogesh P. Wairkar
Richard W. Daniels
Aaron DiAntonio
Published
J Cell Biol. 188: 717-734. 2010
Link
http://jcb-dataviewer.rupress.org/jcb/...
Pubmed
20194640

Grouping

This image is part of a group.
Data Qualifiers
raw, unprocessed data

Sample Preparation

Methods
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
whole mounted tissue

Dimensions

Spatial Axis Image Size Pixel Size
X 2048px ——
Y 2048px ——
*CIL – Cell Image Library accession number. Please use this to reference an image.