For P2X1-inhibition assays, freshly isolated neonatal bovine PMN were pre-treated with NF449 (10 μM, Tocris; purinergic receptor antagonist for P2X1) for 30 min and then co-cultured with C. parvum oocysts (1:2 PMN/oocyst ratio, 3 h, 37°C, 5% CO2). The inhibitor concentration was selected based on previous NET-related studies (15 (link), 40 (link)).
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