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. 2010 Dec 21;104(2):290–299. doi: 10.1038/sj.bjc.6606053

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Copyright © 2011 Cancer Research UK

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AV reovirus possesses a truncated σ1 protein. (A) WT and AV reovirus were purified from L929 murine fibroblasts and examined for the presence of σ1. Through immunoblotting, membranes were probed with reovirus antiserum (left panel) and polyclonal reovirus σ1 N-terminus antiserum (right panel). The N-terminal antibody was raised against σ1 amino acids 1–158 (Duncan and Lee, 1994). (B) Attenuated reovirus was collected from infected HT1080 and PI HTR1 supernatants by high-speed ultracentrifugation and examined by immunoblotting using reovirus antiserum (left panel) and polyclonal σ1 N-terminus antiserum (right panel). Purified WT reovirus was used as a point of reference. (C) In vitro transcription of the AV reovirus S1 gene by T7 polymerase produced a σ1 RNA transcript that was in vitro translated using a rabbit reticulocyte system. Translation of a luciferase transcript and a minus RNA reaction were used as positive and negative controls, respectively. The samples were analysed by 12% SDS–PAGE and subsequent autoradiography.