Under copyright 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. 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.
Rights Holder and Contact
UC Regents
Description
Helix Pomatia Lectin (HPL) (gray) staining is present in diffuse structures that stain with an ER marker, PDI (protein disulfide isomerase) (red) in HeLa cells upon EGF treatment for 4h. The image includes the Golgi (Giantin) (green) 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 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-Giantin, Alexa 594 for anti PDI). 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 4D in J Cell Biol. 189: 843-858. 2010. Images in Fig 4 include CIL#s 13555, 13556, 13557, 13558. Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp) Gill, David J.; Chia, Joanne; Senewiratne, Jamie; Bard, Frederic (2021). CIL:13558, Homo sapiens, epithelial cell, cervical carcinoma. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0959G8R
Type
image
Identifier
ark:/20775/bb67302223
Language
No linguistic content
Subject
Protein folding in endoplasmic reticulum N-acetylgalactosamine binding Golgi organization Protein disulfide isomerase activity HeLa Epithelial cell Cervical carcinoma Endoplasmic reticulum Nucleus Integral to membrane Golgi stack Homo sapiens Cell Image Library Group ID: 6490
If you're wondering about permissions and what you can do with this item, a good starting point is the "rights information" on this page. See our terms of use for more tips.
Share your story
Has Calisphere helped you advance your research, complete a project, or find something meaningful? We'd love to hear about it; please send us a message.
