Fig. 2.

The cytoplasmic domain of the HLA-I heavy chain mediates the interaction between HLA-I and integrin β₄. (A) Schematic representation of the mutant HLA-A2 molecules that have either the extracellular (E-HLA-A2) or the cytoplasmic (C-HLA-A2) domains deleted and replaced with GFP. (B) Endothelial cells were infected with adenoviruses encoding either E-HLA-A2 or C-HLA-A2. More than 90% of the infected cells contained the mutant proteins, as determined by flow cytometric analysis of GFP. Data are representative of 10 independent experiments. (C) Endothelial cells containing E-HLA-A2 or C-HLA-A2 were stimulated with antibodies against either HLA-A2 or GFP, cells were lysed and subjected to immunoprecipitation with Sepharose beads conjugated to antibody against integrin β₄, and samples were analyzed by Western blotting with an antibody against GFP. Endothelial cells from donor EC2 were used in these experiments. For each sample, 10% of the total cell lysate used in the immunoprecipitation was loaded as an input control. Data are representative of three independent experiments.