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. 2009 Jun 30;284(35):23322–23330. doi: 10.1074/jbc.M109.037002

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FIGURE 2. — EL protein forms immunoprecipitable complex during secretion but not after secretion. HEK293 conditioned media were prepared as follows. Lanes 1 and 4 are from cells expressing GFP; lanes 2 and 5 are mixtures of media from cells separately transfected with either EL-myc-His or EL-FLAG; lanes 3, 6, and 7 are media from cells co-transfected with EL-myc-His and EL-FLAG. Western blots (WB) show supernatants (lanes 1–3) and pellets (lanes 4–6) after immunoprecipitation (IP) with mouse anti-Myc from samples as indicated. Lane 7 shows the pellet after immunoprecipitation with mouse anti-FLAG from conditioned media from HEK293 cells co-transfected with EL-myc-His and EL-FLAG. These Western blots used either an antibody against an EL peptide, FLAG tag, or Myc tag for detection. F indicates the full-length monomer size of EL; N indicates the N-terminal domain of EL, and C indicates the C-terminal domain of EL.