TABLE 1

Forskolin-induced increases in the CRE-dependent luciferase-expression (relative light units) and inhibition by 2-methylthio-ADP (2-MeSADP) in CHO Flp-In cells expressing human wild-type and mutant P2Y12-receptor constructs

Cells stably expressing the receptor constructs were transiently transfected with the pCRE-luc vector and stimulated by 1 μM forskolin in the absence or presence of 2-MeSADP for 3.5 h. Means ± S.E. from (n) observations.

CHO Flp-In Cells Expressing the Human P2Y12 Receptor
Wild TypeS101A MutantR256A Mutant
Basal RLU2,230,592 ± 224,149 (106)1,832,489 ± 222,521 (62)1,961,353 ± 181,304 (103)
Forskolin-induced RLU4,281,240 ± 267,455 (190)3,970,596 ± 303,414 (106)3,409,546 ± 197,232* (187)
Inhibition by 1 μM 2-MeSADP, %71.5 ± 1.4 (95)72.6 ± 1.8 (59)64.6 ± 1.7** (89)
Inhibition by 2-MeSADP, log IC50−9.0 ± 0.1−9.2 ± 0.1−8.4 ± 0.1*

  • RLU, relative light units.

  • ↵* , p < 0.05;

  • ↵** , p < 0.01 significant differences from values determined at wild-type receptors (ANOVA followed by the Bonferroni post test or F test).