Fig. 3
From: Adopting the rapamycin trapping assay to track the trafficking of murine MHC class I alleles, H-2Kb
H-2Kb–GFP molecules exit the ER in COPII vesicles. COPII vesicles were generated by an in vitro reaction from MEF cells. Controls for this reaction were the omission of cytosol (lane 3), ATP (lane 4), or addition of dominant-negative Sar1 (T39N; lane 5). COPII vesicles or the corresponding donor microsomal membranes (after the reaction; lane 6–8) were lysed with detergent (in the presence of 10 μM peptide as indicated in lanes 1 and 7). H-2Kb–GFP (upper panel) and Na+/K+ATPase (lower panel) were sequentially immunoprecipitated from the lysates with anti-GFP serum and α6F antibodies, respectively as depicted in panel a. Immunoprecipitates were treated with EndoF1 (lanes 1–7). The band intensities were quantified and plotted in panel (b). One out of three experiments is shown here