CIL:37912, Homo sapiens, epithelial cell
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- Cite This Work
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Gu, Yapeng; Forostyan, Tetyana; Sabbadini, Roger; Rosenblatt, Jody (2021). CIL:37912, Homo sapiens, epithelial cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J02Z146F
- Description
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Apoptotic cells produce and transmit the bioactive lipid sphingosine-1-phosphate (S1P) during extrusion. In this confocal Z-series, an antagonist to the S1P receptor, S1P2, was used to plock signaling. High levels of S1P accumulate in the dying, unextruded cells but not in surrounding cells. The nucleus is in blue, the actin-myosin ring at the basolateral surface is in red and S1P is in green.
- Date Issued
- 2021
- Researchers
- Methods
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Cultured HBE monolayers were treated with 10 µM JTE-013 for 10 min. To induce apoptosis, monolayers were exposed to 1,200 µJ/cm2 UV254 irradiation in a UV series II (Spectroline) and incubated for 2 h before fixation. Cells were fixed with 4% formaldehyde in PBS, permeabilized with 0.5% Triton, blocked and incubated with primary antibody for 1 h (50 µg/ml anti-S1P mAb (LPath Inc.)). Secondary antibody was Alexa Fluor 488 goat anti–mouse IgG. Actin was detected with Alexa Fluor 568–phalloidin (Invitrogen). DNA was detected with 5 µM DRAQ5 (Axxora). Confocal micrographs were obtained using a microscope (TCS SP5; Leica) with a 63x oil lens with an electron-multiplied cooled CCD camera 1,000 x 1,000, 8 x 8 mm2 driven by the IQ software (Andor Technologies). ImageJ was used to stack confocal sections into Z series that were then color combined and reconstructed into 3D image using MetaMorph (GE Healthcare). All images were processed further using ImageJ, Photoshop (Adobe), Illustrator (Adobe), and Quicktime Pro (Quicktime) software. This Z-series corresponds to Fig 4E from J Cell Biol. 2011. 193(4): 667-676
- 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: single-spot confocal microscopy; fluorescence microscopy
Parameter imaged: fluorescence emission
Source of contrast: distribution of a specific protein; distribution of epitope; compartmentalization of stain or label
Visualization methods: Alexa Fluor 488; Alexa Fluor 568; phalloidin; DRAQ5
Processing history: unprocessed raw data
Data qualification: Raw;spatialmeasurements;intensitiesquantitation - Series
- Scientific Name
- Anatomy
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- No linguistic content; Not applicable
- Identifier
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Samplenumber: 37912
- Related Resources
- Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL37912
- J Cell Biol. 2011. 193(4): 667-676. https://www.ncbi.nlm.nih.gov/pubmed/?term=21555463
- 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
- Rights Holder
- 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.
- Digital Object Made Available By
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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