Transfection of NIH 3T3 cells with the p.P54L and p.T111I EFNB1 cDNA constructs. (A) EFNB1 constructs p.P54L and p.T111I were generated by site-directed mutagenesis and transfected into the NIH 3T3 cells. After the transfection, RT-PCR using EFNB1 specific primers was performed. RT-PCR products of the primary NIH 3T3 cells (lane 1), NIH 3T3 cells transfected with wild type, p.P54L, p.T111I EFNB1 cDNA constructs, respectively (lanes 2-4) and Cos-1 cells as a positive control (lane 5) are shown. Size markers are shown in lane M (100 bp DNA ladder, Invitrogen). (B) FACS analysis of the NIH 3T3 cells transfected with wild type and mutant EFNB1 constructs. Grey peaks show maximum of GFP fluorescence in NIH 3T3 transfected cells. Empty peaks show maximum of GFP fluorescence in untransfected control cells.