DOI: 10.22002/ffac5-wc615
title: HeLa cells expressing TRIM5α-YFP

description: HeLa cell line 25 expressing stable TRIM5a-YFP. Correlated light electron microscopy. Concentration of 500 nm blue beads (phosphorex) 1/300. 20 nm gold highly concentrated. Manual blotting with Whatman 40. .  [added 2016-07-13 11:42:02] HeLa cell line 25 expressing stable TRIM5α-YFP. Correlated light electron microscopy. Concentration of 500 nm blue beads (phosphorex) 1/300. 20 nm gold highly concentrated. Manual blotting with Whatman 40. Most of these cells actually displayed mainly autofluorescent puncta
description: <strong>Tilt Series Date:</strong> 2015-03-31</p>
<strong>Data Taken By:</strong> Stephen Carter</p>
<strong>Species / Specimen:</strong> Hela cells expressing TRIM5a-YFP infected Vpr-Ruby HIV-1</p>
<strong>Strain:</strong> </p>
<strong>Tilt Series Settings:</strong>  Single Axis, tilt range: (-60.0°, 60.0°), step: 1°, constant angular increment, dosage: 176.0 eV/Å²,  defocus: -8.0 μm,  magnification: 18000x. </p>
<strong>Microscope:</strong> Caltech Polara</p>
<strong>Acquisition Software:</strong> UCSFTomo</p>
<strong>Upload Method:</strong> reupload_downloaded</p>
<strong>Processing Software Used:</strong> IMOD</p>
<strong>Collaborators and Roles:</strong> Cell line 25 and VSV-G pseudotyped virus Vpr-Ruby provided by Tom Hope</p>
<strong>Purification / Growth Conditions / Treatment:</strong> HeLa cells infected with ZLK 106-06-06-14 HIV-1 GFP Vpr Ruby. 500 µl of media and 500 µl of virus. Virus spinoculated at 16°C at 18 g for 1 hr. Cells were washed with warm media and incubated at 37°C. Cells frozen at later time points, approximately 2.5 hrs after temperature shift.</p>
<strong>Sample Preparation:</strong> Cells grown on gold London finder grids for one day. Grids coated with human fibronectin 0.25 mg/ml and 500 nm red fluorescent F8763 beads.</p>
description: Files available via S3 at s://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/scd2015-03-31-6/rawdata/</p>tomography6.mrc,  Tilt Series (Pixel Size 0.6 nm), 3.4 GB  
        
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        tomography6_part121_13.rec,  Reconstruction (Pixel Size 2.4 nm), 1.1 GB  
        
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        tomography6_orig_orig_orig1_full.rec,  Reconstruction (Pixel Size 0.0 nm), 533.4 MB  
        
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        tomography6_orig_orig_orig_full.rec,  Reconstruction (Pixel Size 0.0 nm), 666.7 MB  
        
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