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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:13566*  Cite 
Description

COP-I gamma1 (green) staining at the Golgi (GM130, a cis-Golgi marker) (red), is redistributed out of the Golgi after EGF treatment of HeLa cells for 4 h. The increase in number and intensity of punctate cytoplasmic COP-I stained structures suggests that the rate of COP-I trafficking is significantly increased. Helix Pomatia Lectin (HPL) (gray) staining is also shown. HPL binds various glycans but the Tn antigen in particular. The Tn antigen refers to terminal α-linked N-acetyl galactosamine residues (GalNAc) linked to Ser or Thr residues. HeLa cells were serum starved overnight in DME (noFBS) and treated with human recombinant EGF (100 ng/ml; Sigma-Aldrich) for 4h. Cells were fixed for 10 min (4% paraformaldehyde) and permeabilized (0.2% Triton X-100). Primary antibody staining followed the manufacturer’s instructions. Cells were subsequently stained for 15–30 min with secondary Alexa Fluor–conjugated antibodies (Alexa 488 for anti-COP-I gamma1, Alexa 594 for anti-GM130). Hoechst (blue) and Alexa 647-conjugated-HPL were added during secondary antibody incubations. Cells were mounted onto glass slides using FluorSave (Merck) and imaged at room temperature using an inverted FluoView confocal microscope (model IX81; Olympus) with fluorescence excitation at 488 nm, 561 nm, and 633 nm and either a 60x objective (U Plan Super Apochromatic [UPLSAPO]; NA 1.35) or 100x objective (UPLSAPO; NA 1.40) using Immersol oil. Microscope coupled with a CCD camera (model FVII). Images were acquired and processed using Olympus FV10-ASW software. Image corresponds to Fig 6B, right panel, in J Cell Biol. 189: 843-858. 2010. Images in Fig 6 include CIL#s 13563, 13564, 13565, 13566, 13567, 13568, 13569, 13570.

Biological Sources
NCBI Organism Classification
Homo sapiens
Cell Type
epithelial cell
cervical carcinoma
Cell Line
HeLa
Cellular Component
Golgi cisterna membrane
nucleus
Golgi-associated vesicle
COPI vesicle coat
Attribution
Names
David J. Gill
Joanne Chia
Jamie Senewiratne
Frederic Bard
Published
J Cell Biol. 189: 843-858. 2010.
Pubmed
PMID: 20498016
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL13566
Archival Resource Key (ARK)
ark:/b7295/w9cil13566
Grouping This image is part of a group.
Sample Preparation
Methods
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 1024px 0.062µm
Y 1024px 0.062µm
Z 1px 0.25µm