CIL:24919, Homo sapiens, epithelial cell, cervical carcinoma
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Huang, Hung-Hsiang; Stanley, Pamela (2021). CIL:24919, Homo sapiens, epithelial cell, cervical carcinoma. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J00000Z0
- Description
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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.
- Date Issued
- 2021
- Researchers
- Technical Details
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Preparation: formaldehyde fixed tissue; detergent permeabilized
Relation to intact cell: dispersed cells in vitro
Item type: recorded image
Imaging mode: fluorescence microscopy
Parameter imaged: fluorescence emission
Source of contrast: distribution of a specific protein; compartmentalization of stain or label
Visualization methods: Alexa Fluor 488; Alexa Fluor 568; 4',6-diamidino-2-phenylindole (DAPI); primary antibody plus labeled secondary antibody; c-MYC peptide tag; HA peptide tag
Data qualification: Processed;spatialmeasurements - Series
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- No linguistic content; Not applicable
- Identifier
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Samplenumber: 24919
- Related Resources
- Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL24919
- J Cell Biol. 187: 967-975. 2009. https://www.ncbi.nlm.nih.gov/pubmed/?term=20026658
- BioStudies (previously JCB DataViewer): https://www.ebi.ac.uk/biostudies/studies/
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- License
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Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International Public License
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- UC Regents
- Copyright
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Under copyright (US)
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.
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.
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Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp)
- Last Modified
2022-08-12