Gill, David J.; Chia, Joanne; Senewiratne, Jamie; Bard, Frederic (2021). CIL:13563, Homo sapiens, epithelial cell, cervical carcinoma. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0Q52NBS

COP-I beta1 (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 beta1, 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 6A, 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.

• 2021

• Bard, Frederic
• Chia, Joanne
• Gill, David J.
• Senewiratne, Jamie

Preparation: formaldehyde fixed tissue; detergent permeabilized
Relation to intact cell: dispersed cells in vitro
Item type: recorded image
Imaging mode: single-spot confocal microscopy
Parameter imaged: fluorescence microscopy
Source of contrast: distribution of a specific protein; distribution of epitope; compartmentalization of stain or label
Visualization methods: Alexa Fluor 647; Alexa Fluor 594; Alexa Fluor 488; primary antibody plus labeled secondary antibody; Helix pomatia agglutinin; Hoechst
Data qualification: Raw;spatialmeasurements;intensitiesquantitation

• Cell Image Library Group ID: 6490

• Homo sapiens

• Cervical carcinoma
• COPI vesicle coat
• Epithelial cell
• Golgi cisterna membrane
• Golgi-associated vesicle
• Nucleus

• COPI coating of Golgi vesicle
• HeLa
• N-acetylgalactosamine binding
• Protein binding
• Retrograde vesicle-mediated transport, Golgi to ER
• Structural molecule activity

• data
• image

• No linguistic content; Not applicable

Samplenumber: 13563

Doi: https://doi.org/10.6075/J0Q52NBS

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL13563

Reference

• J Cell Biol. 189: 843-858. 2010. https://www.ncbi.nlm.nih.gov/pubmed/?term=20498016

Other resource

• BioStudies (previously JCB DataViewer): https://www.ebi.ac.uk/biostudies/studies/

Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International Public License

• UC Regents

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.

Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp)

2022-08-12