Fig. 3.

Results of screening of a library of bioactive compounds (~2000) using three probes: HPr⁺ (50 μM) in the presence of DNA (0.1 mg/mL) as a probe for O₂Superscript>/Superscript> (a and b) and CBA (100 μM, c and d) or Amplex Red (50 μM) in the presence of HRP (0.1 U/mL, e and f) as probes for H₂O₂. dHL60 cells were stimulated with PMA (1 μM) to induce NOX2 activity (a, c, and e). Schemes of oxidation of the probes and the results of screening after normalization (b, d, and f). Plate-to-plate reproducibility data and number of negative/inconclusive/positive hits for each assay. Score “0” (red color) corresponds to negative, “1” (green color) to inconclusive, and “2” (blue color) to positive hits. (This research was originally published in Journal of Biological Chemistry. Zielonka, J., Zielonka, M., VerPlank, L., Cheng, G., Hardy, M., Ouari, O., Ayhan, M. M., Podsiadly, R., Sikora, A., Lambeth, J. D., & Kalyanaraman, B. Mitigation of NADPH oxidase 2 activity as a strategy to inhibit peroxynitrite formation. J. Biol. Chem. 2016; 291:7029–7044. ^© the American Society for Biochemistry and Molecular Biology)