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. 2013 Jan 30;288(12):8627–8635. doi: 10.1074/jbc.M112.441949

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Recombinant progranulin is a homodimer. A, purification of secreted recombinant human progranulin from conditioned media of transfected HEK293T cells is shown. Anti-FLAG Western blot (WB, left) and Coomassie-stained gel (right) of purified progranulin are shown. FT, flow-through. B, recombinant human progranulin migrates at ∼170–180 kDa. Western blot of gel-filtration fractions. Similar results were obtained in two independent experiments. C, cross-linking partially shifts the size of recombinant progranulin from ∼85 to ∼180 kDa. Shown is a Western blot of recombinant human progranulin subjected to cross-linking with disuccinimidyl suberate. D, expressed progranulin-FLAG physically interacts with progranulin-Myc. Co-immunoprecipitation (IP) of conditioned medium from transfected HEK293T cells with anti-FLAG magnetic beads is shown. These results are from a representative experiment that was performed three times with similar results.