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. 2019 Jan 24;104(9):1879–1891. doi: 10.3324/haematol.2018.208819

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Figure 2. — NOX1 is specifically involved in platelet activation by collagen but not thrombin. 1-hydroxy-3-methoxycarbonyl-2, 2, 5, 5-tetramethylpyrrolidine (CMH) was utilized for the detection of oxygen radicals generated by platelets. 10 μg/mL collagen (A) or 0.1 unit/mL thrombin (B) were tested. 10 μM of NoxA1ds abolished the electron paramagnetic resonance (EPR) response measured in the presence of collagen. The scrambled peptide at the same concentration (scNoxA1ds) was used as a negative control. Interestingly, the inhibition of NOX1 by NoxA1ds also inhibited collagen-dependent (C), but not thrombin-dependent (D) platelet aggregation. Aggregation curves for up to 5 minutes (min) are shown, while EPR resonance readings were taken after 10 min of stimulation. Examples of EPR traces and aggregation curves are representative of 3 or more independent experiments. Statistical analysis was performed by one-way ANOVA with Bonferroni post-hoc test for EPR. *P<0.05 compared to resting platelets or t-test for aggregation. *P<0.05 compared to scrambled control. N=3 for (A-D).