The effect of neuraminidase treatment on T1L infectivity in L cells and MEFs.

<p>(<b>A</b>) L cells or (<b>B</b>) MEFs were treated with <i>A. ureafaciens</i> neuraminidase for 1 h, followed by adsorption of T1L at an MOI of 10 or 100 PFU/cell, respectively. Cells were washed twice with PBS, and fresh medium was added. After incubation at 37°C for 20 h, cells were fixed, and reovirus antigen was detected by indirect immunofluorescence. Nuclei were stained with DAPI. The percentage of infected cells in three fields of view per well was determined. The results are expressed as the mean percent infected cells per well in triplicate wells for two independent experiments. Error bars represent standard deviations. (<b>A</b>) n.s., (<b>B</b>) ***, <i>P</i><0.0001, as determined by two-tailed Student's <i>t</i> test. (<b>C</b>) L cells or MEFs were stained with anti-JAM-A antibody followed by Alexa-488 labeled secondary antibody to measure cell-surface JAM-A expression. Fluorescence was detected by flow cytometry. Cells were gated on forward and side scatter and the mean fluorescence intensity (MFI) of Alexa-488 was quantified. Results shown are from a representative of three experiments each done with duplicate samples.