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Detection of LRV with a monocolonal anti-dsRNA (J2) antibody by immunofluorescence microscopy.

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posted on 2013-02-19, 17:18 authored by Haroun Zangger, Catherine Ronet, Chantal Desponds, F. Matthew Kuhlmann, John Robinson, Mary-Anne Hartley, Florence Prevel, Patrik Castiglioni, Francine Pratlong, Patrick Bastien, Norbert Müller, Laurent Parmentier, Nancy Gore Saravia, Stephen M. Beverley, Nicolas Fasel

A. Reference strain analysis (protocol A, see “Material and methods”). Green: dsRNA (J2 Ab). Blue: DAPI (standardized exposure time in all images). B. Phase and immunofluorescent images of Lg M4147 LRVhigh or LRVneg cells were obtained in the presence or absence of J2 antibody (protocol B). C. Quantitative immunofluorescence (protocol B). The fluorescent intensity per cell was assessed using Image J software on Lg M4147 LRVhigh or LRVneg cells following IFM with the J2 antibody. Cells from phase images were identified and the fluorescent intensity average over the area of the cell was recorded. 108–160 cells from 2 distinct fields were measured, and histogram plots were made using Excel software. LRVhigh, no primary antibody (▪, dashed line); LRVhigh with J2 (▪, solid line); LRVneg, no primary antibody (•, dashed line); LRVneg with J2 (•, solid line).

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