CLEC-2–deficient platelets show abolished response to rhodocytin but normal responses to classic agonists. Mice were treated with vehicle or 8 μg/g INU1 and analyzed on day 5. (A) Washed platelets from control (black) or INU1-treated (gray) mice were stimulated with the indicated agonists, and light transmission was recorded on a Fibrintimer 4-channel aggregometer. The results are representative of 6 individual experiments. (B) Flow cytometric analysis of αIIbβ3 integrin activation (binding of JON/A-PE, left panel) and degranulation-dependent P-selectin exposure (right panel) in response to the indicated agonists from control or INU1-treated mice. U46 indicates U46619. Results are mean fluorescent intensities (MFI) ± SD of 6 mice per group. (C) Measurement of released ATP. Washed platelets were incubated for 2 minutes at 37°C with the indicated agonists and fixed. ATP present in the supernatant was measured using a luminometric assay. Results are given as mean ATP concentration (μM) ± SD (n = 6 per group).