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. 2014 May 5;211(5):943–959. doi: 10.1084/jem.20130790

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© 2014 Xiao et al.

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Figure 1. — RGMb binds to PD-L2, but not PD-L1 or other related molecules. (a and b) mRGMb- or mPD-L2–transfected 300 cells, untransfected 300 cells, and PD-1–transfected Jurkat T cells were stained with the indicated Ig fusion proteins (red) or control-Ig (blue) and analyzed by flow cytometry. MFI, mean fluorescence intensity. (c and d) Binding of mPD-L2-Ig or control-Ig to plates coated with mRGMb-HIS, hRGMb-HIS, mRGMa-HIS or mRGMc-HIS, was analyzed by ELISA. (e) Biacore analysis of the interaction of RGMb with PD-L2 and of PD-1 with PD-L2. (f–k) Cell–cell binding of the indicated transfected cells was analyzed by cell conjugation assay. In f, h, and j, the top panels show the FSC-SSC plots and the bottom panels show the corresponding dot plots. g, i, and k show only the dot plots. All data are representative of 2–11 independent experiments.