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. 2013 Aug 8;110(35):14290–14295. doi: 10.1073/pnas.1303380110

Fig. 2.

Fig. 2.

The MHC I glycan is required for EDEM1-dependent HC degradation. (A and B) Nonglycosylated MHC I is degraded via HRD1 and SEL1L. (A) β2m siRNA-treated HeLa cells expressing HLA-A2 S88R were depleted of HRD1 or gp78 and pulse-labeled for 10 min, and MHC I HCs were immunoprecipitated at the indicated times. Graphs represent mean plus SEM (vertical lines) from two independent experiments. (B) MHC I immunoblot in β2m-depleted cells expressing soluble HLA-A2 S88R treated with indicated siRNA. (C and D) EDEM1 is required for degradation of glycosylated, endogenous MHC I. (C) GFP levels in HeLa GFP-HLA-A2wt shβ2m cells treated with EDEM1 or HRD1 siRNA (black line) vs. control siRNA (shaded). (D) β2m-depleted HeLa cells treated with EDEM1 or control siRNA were treated as in A. The graph represents mean plus SEM (vertical lines) from three independent experiments. (E and F) Degradation of nonglycosylated MHC I is EDEM1-independent but requires Herp. (E) MHC I immunoblot in β2m-depleted cells coexpressing soluble HLA-A2 and HLA-A2-S88R treated with indicated siRNA. (F) GFP levels in β2m-depleted cells expressing GFP-HLA-A2wt, S88R, or soluble S88R, treated with EDEM1 or Herp siRNA (black line) vs. control (shaded). Bars represent mean plus 95% CI (black lines) from three independent experiments.