Fig. 4. EDEM accelerates the degradation of misfolded protein in an α1,2-mannosidase-dependent manner. (A) Autoradiogram showing the inhibition of NHK degradation by an α1,2-mannosidase inhibitor kifunensine. Metabolic labelling and immunoprecipitation of transfected HEK 293 cells was performed as described in Figure 3. The position of the predicted Man9 form of the A1AT NHK is shown by *, and that of the Man8 form by **. Intensity of NHK at 0 h chase without kifunensine was arbitrarily set to 1.0, and plotted as in Figure 3A. (B) Degradation of non-glycosylated protein in the ER. pEYFP-ER (Clontech, Palo Alto, CA) was transfected to HEK 293 cells and the intracellular fate of expressed GFP was examined with or without the coexpression of EDEM-HA, as described in A1AT NHK.