EndoH and PNGase were used according to the manufacturer’s instructions (New England Biolabs, Hitchin, UK). Murine monoclonal antibodies were used to detect human HLA-I heavy chain (HC10 hybridoma kindly provided by Professor Hidde Ploegh, Boston Children’s Hospital, USA) and the V5-tag (MCA1360, cloneSV5-Pk1, BioRad, Watford, UK) while rabbit polyclonal antibody specific for actin (A2066, Sigma-Aldritch, Gillingham, UK) was used to check sample loading. Western blots were performed as described previously [27 (link)].
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