{"CIL_CCDB":{"Status":{"Is_public":true,"Deleted":false,"Publish_time":1301284800},"Data_type":{"Still_image":true,"Z_stack":false,"Video":false,"Time_series":false},"CIL":{"Image_files":[{"File_type":"Jpeg","File_path":"13563.jpg","Size":110258,"Mime_type":"image\/jpeg; charset=utf-8"},{"File_type":"OME_tif","File_path":"13563.tif","Size":8500000,"Mime_type":"image\/tif"}],"CORE":{"ATTRIBUTION":{"URLs":[{"Href":"http:\/\/jcb-dataviewer.rupress.org\/jcb\/browse\/2860\/"}],"PUBLISHED":["J Cell Biol. 189: 843-858. 2010."],"Contributors":["David J. Gill","Joanne Chia","Jamie Senewiratne","Frederic Bard"],"PUBMED":["PMID: 20498016"]},"BIOLOGICALPROCESS":[{"onto_name":"retrograde vesicle-mediated transport, Golgi to ER","onto_id":"GO:0006890"},{"onto_name":"COPI coating of Golgi vesicle","onto_id":"GO:0048205"}],"CELLLINE":{"onto_name":"HeLa","onto_id":"MCC:0000219"},"PARAMETERIMAGED":{"onto_name":"fluorescence microscopy","onto_id":"FBbi:00000246"},"IMAGEDESCRIPTION":{"free_text":"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\u2019s instructions. Cells were subsequently stained for 15\u201330 min with secondary Alexa Fluor\u2013conjugated 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."},"ITEMTYPE":{"onto_name":"recorded image","onto_id":"FBbi:00000265"},"CELLULARCOMPONENT":[{"onto_name":"Golgi cisterna membrane","onto_id":"GO:0032580"},{"onto_name":"COPI vesicle coat","onto_id":"GO:0030126"},{"onto_name":"Golgi-associated vesicle","onto_id":"GO:0005798"},{"onto_name":"nucleus","onto_id":"GO:0005634"}],"RELATIONTOINTACTCELL":{"onto_name":"dispersed cells in vitro","onto_id":"FBbi:00000611"},"SOURCEOFCONTRAST":[{"onto_name":"distribution of a specific protein","onto_id":"FBbi:00000597"},{"onto_name":"distribution of epitope","onto_id":"FBbi:00000592"},{"onto_name":"compartmentalization of stain or label","onto_id":"FBbi:00000604"}],"GROUP_ID":"6490","MOLECULARFUNCTION":[{"onto_name":"structural molecule activity","onto_id":"GO:0005198"},{"onto_name":"protein binding","onto_id":"GO:0005515"},{"onto_name":"N-acetylgalactosamine binding","onto_id":"GO:0046871"}],"DATAQUALIFICATION":{"free_text":"RAW;spatialmeasurements;intensitiesquantitation"},"TERMSANDCONDITIONS":{"free_text":"attribution_nc_sa"},"IMAGINGMODE":{"onto_name":"single-spot confocal microscopy","onto_id":"FBbi:00000252"},"DIMENSION":[{"Space":{"Image_size":1024,"Pixel_size":{"unit":"microns","value":0.062},"axis":"X"}},{"Space":{"Image_size":1024,"Pixel_size":{"unit":"microns","value":0.062},"axis":"Y"}},{"Space":{"Image_size":1,"Pixel_size":{"unit":"microns","value":0.25},"axis":"Z"}}],"NCBIORGANISMALCLASSIFICATION":{"onto_name":"Homo sapiens","onto_id":"NCBITaxon:9606"},"PREPARATION":[{"onto_name":"formaldehyde fixed tissue","onto_id":"FBbi:00000010"},{"onto_name":"detergent permeabilized","onto_id":"FBbi:00000262"}],"VISUALIZATIONMETHODS":[{"onto_name":"Alexa Fluor 647","onto_id":"FBbi:00000447"},{"onto_name":"Alexa Fluor 594","onto_id":"FBbi:00000444"},{"onto_name":"Alexa Fluor 488","onto_id":"FBbi:00000440"},{"onto_name":"primary antibody plus labeled secondary antibody","onto_id":"FBbi:00000156"},{"onto_name":"Helix pomatia agglutinin","onto_id":"FBbi:00000429"},{"free_text":"Hoechst"}],"CELLTYPE":[{"onto_name":"epithelial cell","onto_id":"CL:0000066"},{"free_text":"cervical carcinoma"}]}},"Citation":{"Title":"David J. Gill, Joanne Chia, Jamie Senewiratne, Frederic Bard (2011) CIL:13563, Homo sapiens, epithelial cell, cervical carcinoma. CIL. Dataset","ARK":"ark:\/b7295\/w9cil13563","DOI":"doi:10.7295\/W9CIL13563"}}}