FIG 1.

Generation of reoviruses by forward genetics in MDA-MB-231 cells. (A) Triple-negative breast cancer MDA-MB-231 cells were coinfected with T1L, T2J, and T3D and serially passaged 10 or 20 times. Virus isolates were obtained following a plaque assay on L929 cells and sequenced by Illumina next-generation sequencing. (B) Polyacrylamide gel electrophoresis of reovirus parental strains T1L, T2J, and T3D; r1Reovirus (r1); and r2Reovirus (r2). Strains are differentiated by migration patterns of three large (L), three medium (M), and four small (S) gene segments. (C) Percentage of viral isolates with a specific electropherotype following 10 serial passages in MDA-MB-231 cells (n = 44). r1Reovirus accounts for 11% of isolates, while r2Reovirus accounts for 52%. (D) Percentage of viral isolates with a specific electropherotype following 20 serial passages in MDA-MB-231 cells (n = 45). r1Reovirus accounts for 33% of isolates, while r2Reovirus accounts for 27%.