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. 2003 May 19;197(10):1279–1289. doi: 10.1084/jem.20030044

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Copyright © 2003, The Rockefeller University Press

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Figure 4. — UGT2B17 mRNA is transcribed in minor H antigen positive but not minor H antigen negative cells. (A) Northern blot analysis of total RNA from recipient and donor B-LCL, and from HLA-A29⁺ B-LCLs from 4 unrelated donors (designated 1–4). The blot was hybridized with a ³²P-labeled UGT2B17 probe for 16 h at 68°C. Detection of GAPDH mRNA was performed as a control. (B) Cytotoxicity assay for B-LCL by PL8 CTL. The B-LCL from unrelated donors (1–4) are the same lines used for analysis of gene expression in Fig. 3 A. Specific lysis is shown as the mean of triplicate cultures at an E:T ratio of 5:1. (C) Transfection of UGT2B17 cDNA into donor B-LCL restores recognition by PL8 CTL. Donor B-LCL were transfected by electroporation with the full-length construct of UGT2B17, selected for 3 d with puromycin (0.6 μg/ml), and assayed as targets for PL8 CTL. The lysis of UGT2B17-transfected donor B-LCL (triangles), untransfected donor B-LCL (circles), and recipient B-LCL (squares) is shown at various E:T ratios.