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

CIL:13548*  Cite 
Description

After EGF treatment of HeLa cells for 4h, a significant amount of GalNac-T1 staining (red) is apparent in punctate and diffuse cytoplasmic cellular structures that stain positively for Helix Pomatia Lectin (HPL) (green). Anti-Giantin marks the Golgi (gray). The Tn antigen refers to terminal α-linked N-acetyl galactosamine residues (GalNAc) linked to Ser or Thr residues. HPL binds various glycans but the Tn antigen in particular. HeLa cells were serum starved overnight in DME (noFBS) and then 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-GalNac-T1, Alexa 594 for anti-Giantin). Hoechst (blue) and Alexa 647-conjugated-HPL was 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, 543 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 2B in J Cell Biol. 189: 843-858. 2010. Images in Fig 2 include CIL#s 13547, 13548, 13549, 13550.

Biological Sources
NCBI Organism Classification
Homo sapiens
Cell Type
epithelial cell
cervical carcinoma
Cell Line
HeLa
Cellular Component
Golgi stack
integral to membrane
Golgi cisterna membrane
nucleus
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/W9CIL13548
Archival Resource Key (ARK)
ark:/b7295/w9cil13548
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 0.103µm
Y 1024px 0.103µm
Z 1px 5.07µm


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