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. 2022 Dec 16;23(24):16026. doi: 10.3390/ijms232416026

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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In vitro ALOX activity assays. In vitro ALOX activity assays (see Section 4) were carried out using aliquots of the cellular lysates as enzyme source. The reaction products formed from arachidonic acid during a 3 min incubation period were analyzed by RP-HPLC. Conjugated dienes co-eluting with an authentic standard of 12-HETE are detected (panels (A,D,G)). These products were not formed in two different control incubations [no enzyme controls (addition of PBS instead of cellular lysis supernatant to the activity assay), (panels (C,F)) and heat controls, (panels (B,E,H))]. Since we normalized each chromatogram to the highest peak direct comparison of the amounts of products formed by the different enzymes is not possible.