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Description
Helix Pomatia Lectin (HPL) (green) redistribution out of the Golgi (Giantin) (red) in HeLa cells upon EGF stimulation for 4 h is inhibited by co-treatment of Src Kinase Inhibitor, SU6656. The Tn antigen refers to terminal α-linked N-acetyl galactosamine residues (GalNAc) linked to Ser or Thr residues. HPL binds various glycans but the Tn antigen in particular. HeLa cells were serum starved overnight in DME (noFBS) and treated with human recombinant EGF (100 ng/ml; Sigma-Aldrich) and 10 µM SU6656; Merck 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). 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 405 nm, 488 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 3A, bottom right, Src Inhibitor treated/4 hours EGF, in J Cell Biol. 189: 843-858. 2010. Images in Fig 3 include CIL#s 13551, 13552, 13553, 13554. 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:13551, Homo sapiens, epithelial cell, cervical carcinoma. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0GX49BT
Type
image
Identifier
ark:/20775/bb4204597k
Language
No linguistic content
Subject
N-acetylgalactosamine binding Golgi organization Protein O-linked glycosylation via serine Polypeptide N-acetylgalactosaminyltransferase activity Protein O-linked glycosylation via threonine Manganese ion binding HeLa Golgi cisterna membrane Cervical carcinoma Epithelial cell Integral to membrane Golgi stack Nucleus Homo sapiens Cell Image Library Group ID: 6490
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