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

Abstract

<p><b>A.</b> Reference strain analysis (protocol A, see “Material and methods”). Green: dsRNA (J2 Ab). Blue: DAPI (standardized exposure time in all images). <b>B.</b> Phase and immunofluorescent images of <i>Lg</i> M4147 LRV^high or LRV^neg cells were obtained in the presence or absence of J2 antibody (protocol B). <b>C.</b> Quantitative immunofluorescence (protocol B). The fluorescent intensity per cell was assessed using Image J software on <i>Lg</i> M4147 LRV^high or LRV^neg 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. LRV^high, no primary antibody (▪, dashed line); LRV^high with J2 (▪, solid line); LRV^neg, no primary antibody (•, dashed line); LRV^neg with J2 (•, solid line).</p