Utilization of sialic acid as a coreceptor is required for reovirus-induced biliary disease
J. Clin. Invest. Erik S. Barton, et al. 111:1823 doi:10.1172/JCI16303 [Go to this article.]

Figure 3
Genotypic and phenotypic characterization of virus isolated from mice following infection with T3SA– and T3SA+. (a) Viral genomic RNA was extracted from either homogenized livers of infected mice or second-passage lysate stocks. S1 gene segment cDNA’s were amplified using RT-PCR (labeled at right as Primary and Secondary), and restriction enzyme digestion was performed using BstNI. Digested secondary PCR products (labeled at right as 355 bp and 190 bp) were visualized by agarose gel electrophoresis and ethidium bromide staining. Undigested (U) and digested (D) stock virions are shown along with infected livers from days 4, 8, and 12 after inoculation. The lanes labeled M were loaded with 100-bp markers. (b) Homogenized livers obtained 4, 8, and 12 days after inoculation were passaged in L cells for 72 hours. L cell lysate (150 μl) was adsorbed to either L cells or MEL cells (2 × 10⁵) and incubated at 37°C for 24 hours. Viral titers at 0 hours and 24 hours were determined from liver samples obtained from three to four mice by plaque assay using L cells. Viral yields were calculated by dividing titer at 24 hours by titer at 0 hours, and average viral yields for all at 3 days after inoculation are shown. Error bars indicate SEM.