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. 2010 Mar 30;285(23):17564–17573. doi: 10.1074/jbc.M110.111641

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 10. — A Tg-AChE chimera exhibiting functional cholinesterase activity is nevertheless incompetent for secretion. A, 293 cells transfected to express either wild-type Tg (as a positive control), Tg-AChE, or Tg-AChEΔCys were pulse-labeled for 30 min and chased for 3.5 h. The media were collected, and the cells were lysed and immunoprecipitated with anti-Tg before analysis by reducing SDS-PAGE and fluorography. B, specific cholinesterase activity of Tg-AChE and Tg-AChEΔCys (normalized to Western blotting with anti-Tg). C, an independent experiment similar to panel A but analyzed by non-reducing SDS-PAGE and fluorography. Despite that AChE has a smaller molecular mass than ChEL, Tg-AChE and Tg-AChEΔCys run slower (higher), because they are oxidatively immature. Untransfected cells (Control) are shown in the first two lanes of panels A and C. The position of a 205-kDa molecular mass marker is indicated.