FIG 6 .

CHO cells stably expressing hJAM1 support Hom-1 replication. (A) Cells stably expressing full-length hJAM1 (CHO-J) or hJAM1 with the C-terminal sequence deleted (Δ261-299 aa; CHO-T), as well as cells transfected with an empty expression vector (CHO-D), were a gift from T. Dermody. Expression of hJAM1 derivatives in these cells was confirmed by Western blotting with anti-hJAM1 antibodies (Acris Antibodies). (B) Immunofluorescent staining of CHO, CHO-D, CHO-J, and CHO-T cells infected with Hom-1. Cells were mock infected or inoculated with Hom-1 at an MOI of 1, incubated at 37°C for 16 h, and fixed with 4% PFA–0.1% Triton X-100 before being stained with guinea pig anti-Hom-1 VLP and mouse anti-hJAM1 (BV16) antibodies. The bound antibodies were visualized with goat anti-mouse IgG and anti-guinea pig IgG antibodies labeled with the Alexa Fluor 488 and 594 dyes, respectively. (C) Growth curve of Hom-1 in stably transfected CHO cells. CHO cells (n = 10⁷) were inoculated with Hom-1 at an MOI of 1. After 1 h of incubation at 37°C, the cells were washed to remove unadsorbed virus. Next, growth medium was added and the cells were maintained at 37°C for various times. Infected cells were then collected with growth medium, and virus titers in Vero cells were determined with a plaque-forming assay. The data represent the mean titers of two replicate experiments with the standard error shown for each point.