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. 2019 Feb 22;9:2567. doi: 10.1038/s41598-018-37495-6

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Cocaine induces iNOS to regulate prolidase phosphorylation. (A) Cellular lysates of untreated and cocaine-treated HBMECs were analyzed for iNOS expression by immunoblot. (B) Densitometry analysis of iNOS expression relative to actin from three independent experiments. Immunoblot analysis of phospho-prolidase expression. (C) eNOS expression in the cellular lysates of untreated and cocaine-treated HBMECs. (D) Measurement of NO levels in the culture supernatant of HBMECs treated without or with increasing concentrations of cocaine. (E) Effects of iNOS inhibition on NO production. HBMECs were treated with cocaine (25 μM) or iNOS inhibitor (SMT) followed by measurement of NO levels in the culture supernatant. (F) Effects of iNOS inhibition on phosphorylation of prolidase. HBMEC monolayer was treated with cocaine (50 μM) or iNOS inhibitor (SMT) followed by measurement of phospho-prolidase levels. (G) Densitometry analysis of iNOS inhibition on phospho-prolidase levels relative to total prolidase from three independent experiments. (H) Effects of iNOS inhibition on cocaine-induced transmigration of THP-1 monocytes. HBMEC monolayer was treated with cocaine (50 μM) and/or iNOS inhibitor (SMT) followed by addition of THP-1 cells. Data represents the mean ± SEM of at least three determinations. *P < 0.05 represents statistical comparison of untreated and cocaine-treated samples in Panels (B,D,E,H), whereas **P < 0.05 depicts that of cocaine-treated sample with and without iNOS inhibitor in Panels (E,G,H).