American Society of Hematology

Fig. 5.

$Fig. 5. Analysis of the ligand binding function of the vIIbβ3 chimeric receptor. CHO cells stably transfected with vIIbβ3 were stained with the anti-v MoAb 142 (A) or the IIbβ3-specific ligand mimetic antibody PAC1 (B). To analyze PAC1 binding, cells were incubated with the activating antibody anti-LIBS6 and washed and then PAC1 binding was analyzed in the presence (open histogram) and absence (shaded histogram) of the inhibitor Ro43-5054. Stained, untransfected CHO cells are depicted by the dotted histogram. (C) Lysates of CHO cells stably transfected with vIIbβ3were loaded onto a 1-mL GRGDSPK-Sepharose column. The column was washed and then eluted with 1 mmol/L GRGDS peptide (3 mL). Fractions were resolved on a 8% nonreducing polyacrylamide gel, transferred to nitrocellulose, and immunoblotted with MoAb 142. L, lysate; FT, flow through; LW, last wash fraction; CE, eluted fraction.$

Analysis of the ligand binding function of the _v_IIbβ₃ chimeric receptor. CHO cells stably transfected with _v_IIbβ₃ were stained with the anti-_v MoAb 142 (A) or the _IIbβ₃-specific ligand mimetic antibody PAC1 (B). To analyze PAC1 binding, cells were incubated with the activating antibody anti-LIBS6 and washed and then PAC1 binding was analyzed in the presence (open histogram) and absence (shaded histogram) of the inhibitor Ro43-5054. Stained, untransfected CHO cells are depicted by the dotted histogram. (C) Lysates of CHO cells stably transfected with _v_IIbβ₃were loaded onto a 1-mL GRGDSPK-Sepharose column. The column was washed and then eluted with 1 mmol/L GRGDS peptide (3 mL). Fractions were resolved on a 8% nonreducing polyacrylamide gel, transferred to nitrocellulose, and immunoblotted with MoAb 142. L, lysate; FT, flow through; LW, last wash fraction; CE, eluted fraction.

This Feature Is Available To Subscribers Only