FIG 2.

Growth kinetics of rLASV(IGR/S-S) in cultured cells. (A and B) A549 (A) and Vero (B) cells were inoculated with rLASV-WT or rLASV(IGR/S-S) at the indicated MOIs, and viral titers in TCS were determined by plaque assay at the indicated time points. Data represent means ± standard deviations (SD) of results from triplicate samples. Dotted lines indicate the lower limit of detection (33 PFU/ml). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (Student’s t test). (C) Plaque morphologies and sizes of rLASV-WT (top) and rLASV(IGR/S-S) (bottom) on Vero cell monolayers. Data presented are means ± SD of measurements of 25 randomly selected plaques. (D and E) Replicates of A549 (D) and Vero (E) cells infected as described above were evaluated for LASV NP and GP expression by immunofluorescence assay using anti-LASV GP and NP monoclonal antibodies (MAbs), respectively. Representative images of three independent infection experiments are illustrated. Scale bars = 200 μm. (F) Western blot analysis of LASV-GP and NP expression in A549 cells infected with rLASV-WT or rLASV(IGR/S-S) (MOI = 0.1). Numbers below bands correspond to densitometry quantification of each band normalized to actin beta control.