title: Response to Pagano et al.
abstract: In their letter, Pagano et al. appreciate the development of the Nox1, Nox2, and Nox4 triple (3N(-/-)) knockout mouse. They also agree on the view that chemiluminescence assays in general have severe limitations. However, they criticize the fact that the membrane assays in the particular study were restricted to chemiluminescence techniques. Moreover, Pagano et al. got the impression that statements concerning membrane assays of Nox activity in general were made. In addition to a lack of some technical details, Pagano et al. also found the characterization of the 3N(-/-) incomplete and some of the results to be incomprehensible. Although we are grateful for the interest of Pagano et al. in our work, we realized that basically each observation of our study was questioned. This is certainly an excessive rejection of the study in total and fails to appreciate the clear chain of evidences presented. Our work focused on chemiluminescence, and thus, any conclusions are restricted to this technique. Moreover, the 3N(-/-) mice were never developed to study the physiology of Nox enzymes, but rather to validate Nox specificity of NADPH-stimulated chemiluminescence assays. We are convinced that our findings are a valid demonstration that chemiluminescence-based assays in membrane preparations stimulated with NADPH do not measure Nox activity. This conclusion is based on both overexpression studies as well as genetic deficient mouse models. The criticisms of Pagano et al. thus might be justified in some aspects; they, however, cannot disprove the conclusions of our work. Antioxid. Redox Signal. 23, 1247-1249.
authors.affiliation: 1 Institute for Cardiovascular Physiology, Goethe-University Frankfurt , Frankfurt, Germany .
authors.forename: Flávia
authors.initials: F
authors.lastname: Rezende
authors.affiliation: 1 Institute for Cardiovascular Physiology, Goethe-University Frankfurt , Frankfurt, Germany .
authors.forename: Oliver
authors.initials: O
authors.lastname: Löwe
authors.affiliation: 1 Institute for Cardiovascular Physiology, Goethe-University Frankfurt , Frankfurt, Germany .
authors.forename: Valeska
authors.initials: V
authors.lastname: Helfinger
authors.affiliation: 1 Institute for Cardiovascular Physiology, Goethe-University Frankfurt , Frankfurt, Germany .
authors.forename: Kim-Kristin
authors.initials: KK
authors.lastname: Prior
authors.affiliation: 1 Institute for Cardiovascular Physiology, Goethe-University Frankfurt , Frankfurt, Germany .
authors.forename: Maria
authors.initials: M
authors.lastname: Walter
authors.affiliation: 2 Institute for Vascular Signaling, Goethe-University Frankfurt , Frankfurt, Germany .
authors.forename: Sven
authors.initials: S
authors.lastname: Zukunft
authors.affiliation: 2 Institute for Vascular Signaling, Goethe-University Frankfurt , Frankfurt, Germany .
authors.forename: Ingrid
authors.initials: I
authors.lastname: Fleming
authors.affiliation: 3 Excellencecluster Cardiopulmonary System, Justus-Liebig-University Giessen , Giessen, Germany .
authors.forename: Norbert
authors.initials: N
authors.lastname: Weissmann
authors.affiliation: 1 Institute for Cardiovascular Physiology, Goethe-University Frankfurt , Frankfurt, Germany .
authors.forename: Ralf P
authors.initials: RP
authors.lastname: Brandes
authors.affiliation: 1 Institute for Cardiovascular Physiology, Goethe-University Frankfurt , Frankfurt, Germany .
authors.forename: Katrin
authors.initials: K
authors.lastname: Schröder
mesh_terms: D000818:Animals; D008562:Membrane Glycoproteins / Q000378:metabolism; D018345:Mice, Knockout; D019255:NADPH Oxidases / Q000378:metabolism*; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species
chemical_list: D008562:Membrane Glycoproteins; D017382:Reactive Oxygen Species; D019255:NADPH Oxidases