LJ001 oxidizes unsaturated fatty acids in viral membranes.

<p>(<b>A</b>) Equivalent titers of Semliki forest virus (SFV) grown in cholesterol-depleted or control C6/36 mosquito cells were treated with increasing concentrations of LJ001 and their infectivity on target BHK cells determined by immunofluorescence (as in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003297#ppat.1003297.s002" target="_blank">Figure S2</a>). <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003297#s2" target="_blank">Results</a> are presented as % of infection (mean ± SD, n = 3) relative to that obtained in the absence of LJ001 treatment. The IC₅₀ for LJ001's antiviral activity was determined by non-linear regression using GraphPad PRISM (Top = 100%, Bottom = 0%). (<b>B–C</b>) Purified influenza A virus (A/PR/8/34 H1N1) was treated with 5 µM of LJ001, or control LJ025, and exposed to light for 1 h. The total lipid content was extracted and the viral lipidome analyzed by high-resolution LC-MS (see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003297#ppat.1003297.s003" target="_blank">Figure S3</a>). (<b>B</b>) Relative molar concentration of the major phospholipid species present in the viral lipidome. (<b>C</b>) The amount of peroxidized phosphatidylcholine (PC) species, presented as fold-increase in LJ001- over LJ025-treated samples. Similar results were obtained in two independent experiments with two technical replicates each. PE: Phosphatidylethanolamine, PS: Phosphatidylserine, SM: Sphingomyelin, (OO)ePC: oxidized (hydroperoxide) ether PC, (OO)PC oxidized (hydroperoxide) PC. (<b>D</b>) Liposomes (150 µg in 1 ml) were treated with LJ001 (10 µM), or control LJ025, and exposed to light for 1 h. After de-esterification, fatty acids were extracted, and the amount of 9-hydroxy-10E,12Z-octadecadienoic acid (9-HODE) and 13-hydroxy-9Z,11E-octadecadienoic acid (13-HODE) was determined by LC-MS/MS. Data represents the mean ± SD of triplicates. ****: p<0.0001, LJ001 vs LJ025, Two-way ANOVA, Bonferroni post-test using GraphPad PRISM.