Detection and characterization of PAFR stably transfected in CHO-αIIbβ3 cells.

(A) Cell extracts were subjected to Western blotting analysis using a rabbit polyclonal anti-PAFR. The arrow points to a band of the expected size of PAFR that is present in human platelets and CHO-αIIbβ3-PAFR cells, but absent in CHO-αIIbβ3 cells transfected with the void plasmid. (B) and (C) depict the effects of PAF in increasing the intracellular cytosolic levels of free calcium and intracellular pH, and the effect of a PAF antagonist in preventing both effects. Measurement of intracellular calcium and pH was carried out fluorimetrically as described in “Materials and methods.” The data are representative of at least 6 observations made in different cell preparations.