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. 2021 Oct 18;29(3):670–686. doi: 10.1038/s41418-021-00883-z

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© The Author(s) 2021

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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Fig. 3 — Global lipidomics analysis of Ms4a15 OE, control cells treated with Tgn^short (3 h), control cells treated with Tgn^long (16 h), and untreated control cells. A Dendrogram indicating separation of untreated Ms4a15 OE and Tgn^long treated control cells from untreated and Tgn^short treated control cells by hierarchical cluster analysis. Similarly regulated lipid species from Ms4a15 OE and Tgn^long were extracted and plotted in the heatmap. B Lipid abundance heatmap showing z-score profiles of species similarly downregulated in both Ms4a15 OE and Tgn^long (group I), exclusively upregulated in Ms4a15 OE (group II), and similarly upregulated in Ms4a15 OE and Tgn^long (group III). Sample colors correspond to A. (n = 3, Wilcoxon–Mann–Whitney-Test, BH corrected). C Modulated lipid classes in groups I–III by LIPID MAPS Structure Database. GP glycerophospholipid, FA fatty acid, ST sterol Lipid, SP sphingolipid, GL glycerolipid. Ether GPs and ester GPs are in dark colors. D Free fatty acid fold change in Ms4a15 OE and Tgn^long compared to untreated control. SFAs saturated fatty acids, MUFAs monounsaturated fatty acids, PUFAs polyunsaturated fatty acids. Significant p-values of two-way t-test comparisons versus control are shown. E Kendrick plot of significantly modulated diacylglycerophospho-ethanolamine (PE) and -choline (PC) ester phospholipids. All species have a referenced Kendrick mass-defect (RKMD) value of 0 (saturated chains) or a negative integer (number of unsaturated bonds). Dot sizes indicate absolute values of log₂(mean Ms4a15 OE/mean control) (n = 3). p < 0.05, Wilcoxon–Mann–Whitney-Test, BH corrected). F Model structures of diacyl (esters), plasmanyl (ethers) and plasmenyl (vinyl-ethers). The latter are also termed plasmalogens. G Kendrick plot of significantly modulated ether GPs (PE and PC). Dot sizes indicate summed peak intensity. For given species isomeric plasmalogens are validated by acidic hydrolysis (see Supplementary Table 2) and (H). ‘PC e’ or ‘PE e’ represent the respective ether species of PC or PE, n = 3, p < 0.05, Wilcoxon–Mann–Whitney-Test, BH corrected). H Acidic hydrolysis abundance illustrated for one ester (top), one alkyl-ether (middle) and one vinyl-ether GP (bottom). I Summed intensities for all detected GP show a slight reduction of ester GPs as well as enrichment in ether GP for Ms4a15 OE and Tgn^long. Data shown represent mean ± SD of n = 3 technical replicates.