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Fig. 8. ER mannosidase I inhibitors block ERAD of wild-type Shaker protein. (A) Shaker-expressing cells were pulsed for 10 minutes and either harvested immediately or chased for 48 hours. Cells were treated with BFA/NOC and no additional drug; BFA/NOC and dNJ (1 mM); BFA/NOC and dMJ (1 mM); BFA/NOC, dNJ and dMJ; BFA/NOC, dNJ and KIF (10 µM); or BFA/NOC, dNJ and SWN (1 mM). A representative fluorograph is shown. Open arrowhead indicates the position of the core glycosylated, immature form of the Shaker protein. (B) The amount of core glycosylated Shaker protein remaining at 48 hours of chase was quantified by densitometry and expressed relative to the amount present at 0 time of chase (100%). The number of experiments for each condition (n) is shown in parentheses on the appropriate bar. P<0.001 compared with all other treatments except dNJ+SWN; Student's t-test. Data are presented as mean±s.e.m. except for the dMJ sample, where n=2. For the dMJ sample, the mean value is shown.

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