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. 2002 Apr 2;21(7):1754–1763. doi: 10.1093/emboj/21.7.1754

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Copyright © 2002 European Molecular Biology Organization

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graphic file with name cdf178f6a.jpg — Fig. 6. Tyrosine phosphorylated dimers of STAT1 interact directly with importin-α5. (A) Recombinant (rec) wtSTAT1 was expressed in TKX1 bacteria either unphosphorylated (–) or tyrosine phosphorylated (+) by the Elk kinase. Purified rec STAT1 was incubated with GST (lanes 3 and 4) or GST–importin-α5 (lane 5 and 6) bound to glutathione beads. The binding of rec STAT1 was detected by western blots performed with anti-STAT1 phosphotyrosine antibody (anti-STAT1 pY) (upper panel) or anti-STAT1 antibody (lower panel). (B) Recombinant (rec) tyrosine-phosphorylated wtSTAT1 or STAT1 SH2^–(R602L) was purified from TKX1 bacteria expressing Elk kinase and evaluated in an importin-α5 binding assay. The binding of the rec STAT1 proteins was detected by western blot performed with anti-STAT1 phosphotyrosine antibody (anti-STAT1 pY).

graphic file with name cdf178f6b.jpg — Fig. 6. Tyrosine phosphorylated dimers of STAT1 interact directly with importin-α5. (A) Recombinant (rec) wtSTAT1 was expressed in TKX1 bacteria either unphosphorylated (–) or tyrosine phosphorylated (+) by the Elk kinase. Purified rec STAT1 was incubated with GST (lanes 3 and 4) or GST–importin-α5 (lane 5 and 6) bound to glutathione beads. The binding of rec STAT1 was detected by western blots performed with anti-STAT1 phosphotyrosine antibody (anti-STAT1 pY) (upper panel) or anti-STAT1 antibody (lower panel). (B) Recombinant (rec) tyrosine-phosphorylated wtSTAT1 or STAT1 SH2^–(R602L) was purified from TKX1 bacteria expressing Elk kinase and evaluated in an importin-α5 binding assay. The binding of the rec STAT1 proteins was detected by western blot performed with anti-STAT1 phosphotyrosine antibody (anti-STAT1 pY).