The identification, confirmation, and expression of non-Gal porcine aortic endothelial cell (PAEC) antigens. (A) Flow cytometry of IgG binding to pRETRO-infected HEK cells for individual non-Gal antigens. (B) Flow cytometry showing IgG binding to G418-resistant pcDNA3.1/V5-His-TOPO transfected cell lines expressing individual non-Gal antigens. (C) Reverse-transcriptase polymerase chain reaction analysis of non-Gal gene expression. RNA samples: lane 1, pig heart; lane 2, GTKO PAECs; lanes 3 to 5, independent G418-resistant HEK transformants; lane 6, untransformed HEK cells; lane 7, a flow cytometry negative HEK cell line; and lanes 8 to 10, the same as lanes 3–5 but without reverse transcriptase. Non-Gal antigen expression: (a, g, and m) CD9; (b, h, and n) PROCR; (c, i, and o) CD46; (d, j, and p) CD59; (e, k, and q) ANXA2; and (f, l, and r) B4GALNT2.