Extended Data Fig. 10: Characterization of the anti-R9AP monoclonal antibody 5E9 and its effects on blocking EBV infection.
From: R9AP is a common receptor for EBV infection in epithelial cells and B cells
a, Complementarity determining regions (CDR) details for the anti-R9AP monoclonal antibody 5E9 antibody. b, ELISA binding for 5E9 and control antibody to R9AP1-210. The A450 signals were from triplicate wells and the mean signal were presented. c, BLI binding assay for 5E9 to R9AP1-210. 5E9 were captured onto ProA biosensors and assayed for binding to R9AP1-210. d, WB for 5E9 binding to R9AP in comparison to commercial antibody. HEK-293T cells were transfected with FLAG-R9AP. Cells were lysed and immunoprecipitated with 5E9 or anti-R9AP antibody (Cat #HPA049791, Sigma). e, Primary B cells were pretreated with anti-R9AP monoclonal antibody (5E9), then infected with EBV. The proportion of EBERs-positive cells were independently evaluated by three pathologists. Each pathologist counted 3 representative high-power fields (×40 objective) per sample, with approximately 100 cells/field, and obtained a mean value. Bars represent proportion of EBERs-positive cells. Data are mean ± S.E.M. and representative of two independent experiments (n = 3). f-j, Akata, Raji, HNE1, AGS and HEK-293T cells were pretreated with anti-R9AP monoclonal antibody (5E9), then infected with EBV. EBV infection efficiency was analyzed by flow cytometry. Data are representative of two (b-d) or three independent experiments (f-j).
