Extended Data Fig. 6: R9AP, EPHA2, and NRP1 are not redundant for EBV infection of epithelial cells.

a-c, Cell-based EBV fusion assay by co-culture of sgVector transfected HNE1 cells, EPHA2 knockout HNE1 cells, R9AP overexpressed while EPHA2 knockout HNE1 cells or EPHA2 rescued EPHA2 knockout HNE1 cells, and HEK-293T cells transfected with gH/gL/gB (a). EBV infection efficiency was analyzed by flow cytometry (c) and quantified (b) in the above-indicated HNE1 cells. d-f, Cell-based EBV fusion assay by co-culture of sgVector transfected HNE1 cells, R9AP knockout HNE1 cells, EPHA2 overexpressed while R9AP knockout HNE1 cells or R9AP rescued R9AP knockout HNE1 cells, and HEK-293T cells transfected with gH/gL/gB (d). EBV infection efficiency was analyzed by flow cytometry (f) and quantified (e) in the above-indicated HNE1 cells. g, HEK-293T cells were transfected with Myc-R9AP, EPHA2 together with FLAG-gH/gL or EV, lysed, and immunoprecipitated with antibody against FLAG as indicated IPx1:FLAG. The immunoprecipitated proteins were eluted by FLAG peptide and re-immunoprecipitated with antibody against Myc as indicated IPx2: Myc, followed by WB analysis with indicated antibodies. Data are representative of two independent experiments. h-j, Cell-based EBV fusion assay by co-culture of sgVector transfected HNE1 cells, R9AP knockout HNE1 cells, NRP1 overexpressed while R9AP knockout HNE1 cells or R9AP rescued R9AP knockout HNE1 cells, and HEK-293T cells transfected with gH/gL/gB (h). EBV infection efficiency was analyzed by flow cytometry (j) and quantified (i) in the above-indicated HNE1 cells. Two independent experiments in triplicates (n = 6) and mean and S.E.M. of those n = 6 were used (a, d, h, i), or three independent experiments in triplicates (n = 9) and mean and S.E.M. of those n = 9 were used (b, e). One-way ANOVA was carried out with Tukey’s correction for multiple comparisons. Data are representative of three (c, f) or two (j) independent experiments.

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