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Description
COP-I gamma1 (green) staining at the Golgi (GM130, a cis-Golgi marker) (red) in unstimulated HeLa cells. 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). 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 gamma1, 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 6B, left panel, in J Cell Biol. 189: 843-858. 2010. Images in Fig 6 include CIL#s 13563, 13564, 13565, 13566, 13567, 13568, 13569, 13570. 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:13565, Homo sapiens, epithelial cell, cervical carcinoma. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0B856TF
Type
image
Identifier
ark:/20775/bb7242174w
Language
No linguistic content
Subject
COPI coating of Golgi vesicle N-acetylgalactosamine binding Retrograde vesicle-mediated transport, Golgi to ER Protein binding HeLa Structural molecule activity Golgi-associated vesicle Nucleus COPI vesicle coat Golgi cisterna membrane Epithelial cell Cervical carcinoma Homo sapiens Cell Image Library Group ID: 6490
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