HLA-C cell surface expression was measured in transfected or transduced cells using alexa fluor 647 anti-human HLA-A, B, C antibody (clone W6/32, BioLegend, 311414, 1:20) or anti-HA.11 antibody (clone 16B12, Covance, MMS-101R, 1:250) along with allophycocyanin goat anti-mouse Ig (BD Biosciences, 550826, 1:50). HLA-C or HA staining was determined on GFP+ cells. For intracellular staining, cells were fixed and permeabilized using BD Cytofix/Cytoperm Fixation/Permeabilization kit (BD Biosciences), followed by staining using phycoerythrin anti-human HLA-A, B, C antibody (clone W6/32, BioLegend, 311406, 1:20) or anti-HA.11 antibody (clone 16B12, Covance, MMS-101R, 1:250) along with allophycocyanin goat anti-mouse Ig (BD Biosciences, 550826, 1:50). HLA staining using the same antibodies on non-transfected cells was used as a negative control. Data obtained were analysed using the FlowJo software (Tree Star).
Kaur G., Gras S., Mobbs J.I., Vivian J.P., Cortes A., Barber T., Kuttikkatte S.B., Jensen L.T., Attfield K.E., Dendrou C.A., Carrington M., McVean G., Purcell A.W., Rossjohn J, & Fugger L. (2017). Structural and regulatory diversity shape HLA-C protein expression levels. Nature Communications, 8, 15924.
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