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
GalNac-T1 (green) and ERGIC53 (ER-to-Golgi intermediate compartment) (red) colocalize in unstimulated HeLa cells moderately only at the Golgi apparatus. Helix Pomatia Lectin (HPL) (gray) staining is included. 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). 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-GalNac-T1, Alexa 594 for ERGIC53). 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, 543 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 4A 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:13555, Homo sapiens, epithelial cell, cervical carcinoma. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0XG9PT3
Type
image
Identifier
ark:/20775/bb8504980j
Language
No linguistic content
Subject
Protein O-linked glycosylation via serine Manganese ion binding Protein O-linked glycosylation via threonine Polypeptide N-acetylgalactosaminyltransferase activity HeLa N-acetylgalactosamine binding Golgi cisterna membrane Cervical carcinoma Endoplasmic reticulum-Golgi intermediate compartment Golgi stack Epithelial cell Nucleus Integral to membrane 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.
