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. 2017 Sep 8;7:10981. doi: 10.1038/s41598-017-11496-3

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© The Author(s) 2017

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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LPAT activity assay by double choice of substrate on polarized BMDMs. Microsomes were used to test BMDM - LPAT activity using a mixture of six lysophospholipids and eight acyl-CoA esters as described in Materials and Methods. The total area ratio of all newly formed phospholipids is expressed in the radar graphic (A). The specific production of AA-anchored (B) or DHA-anchored phospholipids (C) is represented for polarized BMDMs. The results indicate the area ratio for the incorporation of fatty acid residues in polar groups by internal deuterated standard. Data are representative for BMDMs, 24 h adherent (M0), primed with IFN-γ for 24 h (M1) or with IL-4 + IL-13 for 24 h (M2). All 48 possible products were quantified using HPLC-MS/MS. The Y-axis represents the area ratio (Log2) of newly formed phospholipids. Two independent microsomal preparations were tested in duplicate and the data shown represent the means ± s.e.m. * p < 0.05 comparing M1 or M2 to M0; ^# p < 0.05 for M2 compared to M1.