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Figure 4 | Retrovirology

Figure 4

From: Amino acid residues that are important for Hyal2 function as a receptor for jaagsiekte sheep retrovirus

Figure 4

Cell-surface mouse Hyal2 expression levels are at least as high as those of human Hyal2 following expression plasmid transfection. Plasmids encoding amino-terminal Flag-tagged mouse or human Hyal2, or an empty plasmid as a control, were cotransfected with pSV2neo at a 10:1 ratio into NIH 3T3 cells. One day after transfection the cells were treated with trypsin and seeded in medium containing G418. Two weeks later, cell-surface Flag-tagged protein expression was measured by FACS after incubation of the cells with an anti-Flag mouse monoclonal primary antibody followed by a goat anti-mouse IgG secondary antibody. Green indicates cells transfected with control plasmid, red indicates cells transfected with Flag-tagged human Hyal2, and orange indicates cells transfected with Flag-tagged mouse Hyal2. Control analyses included Flag-tagged human Hyal2-transfected cells that were not incubated with antibody (purple) or with secondary antibody only (blue). The percentages of cells in the indicated gate were determined and are shown in the inset. The entire experiment was repeated and the repeat values are shown in parentheses.

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