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Mediators of Inflammation
Volume 2014, Article ID 767061, 11 pages
http://dx.doi.org/10.1155/2014/767061
Research Article

Potent Anti-Inflammatory Activity of Pyrenocine A Isolated from the Marine-Derived Fungus Penicillium paxilli Ma(G)K

1Departamento de Ciências Biológicas, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista “Júlio de Mesquita Filho”, 14801-902 Araraquara, SP, Brazil
2Faculdade de Medicina Ribeirão Preto , Universidade de São Paulo, 14049-900 Ribeirão Preto, SP, Brazil
3Instituto de Química de São Carlos, Universidade de São Paulo, CP 780, 13560-970 São Carlos, SP, Brazil
4Departamento de Bioquímica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista “Júlio de Mesquita Filho”, 13506-900 Rio Claro, SP, Brazil

Received 15 August 2013; Revised 12 November 2013; Accepted 19 November 2013; Published 19 January 2014

Academic Editor: Chiara De Luca

Copyright © 2014 Thaís Regina Toledo et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplemental Figure 1A: Cell Viability Assay. (A) Treated during 18h with pyrenocine A (3.75-0.11 µM) without LPS. (B) Pretreatment procedure: cells were pretreated with different concentrations of pyrenocine A (3.75-0.11 µM) for 2 h and then stimulated with LPS (1 µg/mL) during 18h. (C) Post-treatment procedure: cells were stimulated with LPS (1 µg/mL) for 2 h and then added pyrenocine A (3.75-0.11 µM) during 18h. After the treatment, the Alamar Blue reagent was added in the same point-time with LPS and after 18 h the absorbance was read on a spectrophotometer at 570 nm, using 600 nm as a reference wavelength (normalized to the 600 nm value).

Supplemental Figure 1B. Cell death by pyrenocine A by AnexinV/PI. Cells were treated during 18h with pyrenocine A (3.75-0.94 µM) without LPS. Pretreatment procedure: cells were pretreated with different concentrations of pyrenocine A (3.75-0.94 µM) for 2 h and then incubated with LPS (1 µg/mL) during 18h. Post- treatment procedure: cells were stimulated with LPS (1 µg/mL) for 2 h and then added pyrenocine A during 18h (3.75-0.94 µM). Control: cells untreated (without pyrenocine A or LPS). After the treatment the cells were harvested, stained for Annexin V/PI and acquired on FACSCanto machine and analyzed with FCS Express Software.

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