PDGFRα expression does not increase HCMV gB and gH/gL-induced cell-cell fusion.

Abstract

<p><b>A, </b><b>B, </b><b>C, </b><b>and E</b>) Effector cells were established by transducing <b>A & B</b>) ARPE-19 cells, <b>C</b>) HeLa cells or <b>E</b>) rat-RPE cells with Ad vectors expressing HCMV gH, gL and gB and Ad-tet-trans. Target cells were established by transducing other cells of the same kind with <b>A</b>) Ad-tet-trans vector alone or <b>B, C, and E</b>) Ad-PDGFRα and Ad-tet-trans. <b>D</b>) HeLa effector cells were transduced with Ad vectors expressing HSV glycoproteins gB, gH, gL, and gD and HeLa target cells established by transducing cells with an Ad vector expressing nectin-1. <b>F</b>) rat-RPE effector cells were established by transduction with Ad vectors expressing HSV glycoproteins gB, gH, gL, and gD and rat-RPE target cells were established by transducing cells with an Ad vector expressing nectin-1. In every case, effector cells and target cells were trypsinized then replated in equal numbers in small dishes for 24–36 hr. To assess cell-cell fusion, cell monolayers were fixed and stained with syto-green dye (Invitrogen) and fusion was quantified from micrographic images by counting the number of nuclei involved in syncytia formation divided by the total number of nuclei. The percent of fused cells was calculated by counting fused cells versus unfused cells in 3 fields from 3 separate dishes from 2 experiments and involving over 500 cells and standard deviations indicated.</p