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. 2014 Jun 27;289(32):22140–22150. doi: 10.1074/jbc.M114.560938

FIGURE 2.

FIGURE 2.

Structure of sgp130-E10 and its interaction with Hyper-IL-6. A, schematic overview of sgp130-E10 in complex with IL-6/IL-6R. B, immunoprecipitation with conditioned medium from HEK-293 cell cultures transiently transfected with plasmids coding for sgp130-E10Myc-His or Hyper-IL-6Fc. Supernatants were harvested 2 days after transfection followed by immunoprecipitations (IP) using protein A-agarose beads and Western blot analysis using primary antibodies directed against the Myc (precipitated antibody from cell culture supernatant in unknown concentration) or human Fc tags (0.4 μg/ml). Three independent experiments were performed. A representative experiment is shown. C, immunoprecipitation with conditioned medium from HEK-293 cell cultures transiently transfected with plasmids coding for sgp130-E10Fc or sgp130Fc. Supernatants were harvested 2 days after transfection followed by immunoprecipitations using protein A-agarose beads and Western blot analysis using primary antibodies directed against the human Fc tag (0.4 μg/ml) or IL-6 (2.5 μg/ml). A representative experiment out of three independent experiments is shown.