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

CIL:24919*  Cite 
Description

HeLa cells expressing Myc-GnT1IP-L (Myc-tagged GlcNAcT-1 inhibitory protein, long transcript, membrane-anchored) (red) and GlcNAcT-1-HA (green) were treated with cycloheximide and brefeldin A to cause retrograde transport of Golgi enzymes into the ER. Both proteins had punctate costaining around the nucleus, typical of ERGIC localization, in addition to reticular costaining of the ER after BFA treatment. Transiently expressed Myc-GnT1IP-L is normally primarily localized in the Golgi and ERGIC and partially in the ER (shown in Fig 4; J Cell Biol. 190: 893-910, 2010.). GlcNAcT-1 is normally a resident of the medial Golgi. Transfected HeLa cells were fixed in 3% paraformaldehyde, blocked with 0.2% Triton X-100, 1% FBS, 0.5% BSA in PBS with Ca[2+] and Mg[2+], followed by incubation in primary antibodies (anti-Myc mouse mAb, 9E10 (Covance) and anti–HA mouse mAb, HA.11 (Covance)) and secondary antibodies (Alexa 568 and Alexa 488) and DAPI (blue) to stain nuclei. Cells were mounted using Fluoromount (SouthernBiotech). Immunofluorescent images were digitally acquired on an inverted microscope (Axiovert 200M; Carl Zeiss, Inc.) coupled to a 12-bit cooled charge-coupled device camera (AxioCam MRm Rev. 3; Carl Zeiss, Inc.) controlled by Axiovision software (AxioVs40, version 4.7.2.0; Carl Zeiss, Inc.), using a 100x/1.3 NA oil immersion objective (EC Plan-NeoFluar; Carl Zeiss, Inc.), and saved as tiff files (1388 × 1040, 8 bit). All pictures were treated identically using Adobe Photoshop to remove background and adjust contrast. This image corresponds to Figure 5E, middle panel (merge) in J Cell Biol. 190: 893-910, 2010. Images in Fig 5 include CIL#s 13658, 24919, 24920.

Biological Sources
NCBI Organism Classification
Homo sapiens
Cell Type
epithelial cell
cervical carcinoma
Cell Line
HeLa
Cellular Component
Golgi apparatus
integral to membrane
Golgi membrane
endoplasmic reticulum
endoplasmic reticulum-Golgi intermediate compartment
Attribution
Names
Hung-Hsiang Huang
Pamela Stanley
Published
J Cell Biol. 190: 893-910. 2010.
Pubmed
20805325
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL24919
Archival Resource Key (ARK)
ark:/b7295/w9cil24919
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 1388px 169.333µm
Y 1040px 169.333µm