Computational protocol: Regulation of the subcellular distribution of key cellular RNA-processing factors during permissive human cytomegalovirus infection

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Protocol publication

[…] Protein concentrations were determined using a BCA Protein Assay kit (Pierce) according to the manufacturer's instructions. Absorbance measurements were quantified using SoftMax Pro software (Molecular Devices). [...] Probed cells were examined by IFA using a Bio-Rad MRC 1024 confocal laser-scanning microscope [Center for Microscopy and Image Analysis, George Washington University (GWU) Washington, DC, USA] as described previously (; ; ) or using a Zeiss LSM 510 confocal microscope [Children's Research Institute (CRI), Intellectual and Developmental Disabilities Research Center, Washington, DC, USA]. Images were acquired by sequential excitation at 488, 561 and 633 nm as appropriate using 40× (NA=1.3), 63× (NA=1.4) or 100× [NA=1.35 (GWU) or NA=1.4 (CRI)] oil objectives. Post-acquisition processing and image assembly were performed using ImageJ software (National Institutes of Health, MD, USA), Zeiss ZEN 2008, Adobe Photoshop (v.7.0) and Microsoft PowerPoint 2003. […]

Pipeline specifications

Software tools SoftMax Pro, ImageJ
Applications Laser scanning microscopy, Microscopic phenotype analysis
Organisms Human betaherpesvirus 5
Diseases Cytomegalovirus Infections