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. 2002 Jun;76(12):6364–6369. doi: 10.1128/JVI.76.12.6364-6369.2002

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Copyright © 2002, American Society for Microbiology

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FIG. 1. — SMN and NS2 interact in vitro. (A) Western blot analysis of recombinant GST-tagged NS2 captured by polyhistidine (6-His)-tagged human SMN immobilized on histidine binding resin. The blot was developed using anti-GST monoclonal antibody. (B) BIA of the SMN/NS2 complex. Bound GST monoclonal antibody (shift B), NS2 (shift D), and wild-type SMN (shift F) are indicated. Vertical shifts in the horizontal baseline represent positive interactions as response units (RU) and are indicated by arrows (shifts B, D, and F). hTra2β1 protein served as a negative control and does not bind to NS2 (E). (C) SMN binds to the N-terminal region common to NS1 and NS2. GST-tagged NS variants (NS1, NS2, amino acids 339 to 792, and the NS1 and -2 common domain) were captured using an anti-GST antibody and were then pulsed with wild-type SMN. GST-tagged SMN exon 4 was used as a negative control. Values represent binding of SMN constructs above a background control and the mean of three separate experiments.

FIG. 1. — SMN and NS2 interact in vitro. (A) Western blot analysis of recombinant GST-tagged NS2 captured by polyhistidine (6-His)-tagged human SMN immobilized on histidine binding resin. The blot was developed using anti-GST monoclonal antibody. (B) BIA of the SMN/NS2 complex. Bound GST monoclonal antibody (shift B), NS2 (shift D), and wild-type SMN (shift F) are indicated. Vertical shifts in the horizontal baseline represent positive interactions as response units (RU) and are indicated by arrows (shifts B, D, and F). hTra2β1 protein served as a negative control and does not bind to NS2 (E). (C) SMN binds to the N-terminal region common to NS1 and NS2. GST-tagged NS variants (NS1, NS2, amino acids 339 to 792, and the NS1 and -2 common domain) were captured using an anti-GST antibody and were then pulsed with wild-type SMN. GST-tagged SMN exon 4 was used as a negative control. Values represent binding of SMN constructs above a background control and the mean of three separate experiments.