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. 2022 Jun 22;12:10533. doi: 10.1038/s41598-022-14690-0

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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CERS2 and FADS3 Knockouts: (A) CERS2 KO Volcano plot: Lipid species of the CERS2 KO cell line (n = 6) and the AAVS1 control (n = 3) were compared. P-values of t-test without correction for multiple testing are displayed on the y-axis, fold changes of means are shown on the x-axis. Points with additional outlines indicate lipids significant after correction for multiple testing (Benjamini–Hochberg, 365 lipid species or 29.7% of all species). Sphingolipid species are labeled by their species names. (B) CERS2 KO sphingolipid length distribution: Lipid species have been standardized to the lipid class and summed up by their total number of carbon atoms. Values for individual samples are shown by points and means are indicated by bar plots. Error bars correspond to standard deviations. P-values have been adjusted for the total number length features in all classes (n = 168) and are encoded as follows: * for q < 0.05, ** for q < 0.01, *** for q < 0.01,**** for q < 0.0001. Only lipid species were used, which had at least 2 valid measurements in each of the two cell lines (n = 1228). The CERS2 KO clones A10 and B11 are displayed, which were created by the deletion method. (C) FADS3 KO Volcano plot: Lipid species of the FADS3 KO cell line (n = 3) and the AAVS1 control (n = 3) were compared. P-values of t-test without correction for multiple testing are displayed on the y-axis, fold changes of means are shown on the x-axis. Points with additional outlines indicate nominal significant lipids (87 lipid species or 8.2% of all species). Sphingolipid species are labeled by their species names. (D) FADS3 KO sphingolipid double bond distribution: Lipid species have been standardized to the lipid class and summed up by their total number of double bonds. Values for individual samples are shown by points and means are indicated by bar plots. Error bars correspond to standard deviations. p-values have been adjusted for the total number db features in all classes (n = 113) and are encoded as follows: * for q < 0.05, ** for q < 0.01, *** for q < 0.01,**** for q < 0.0001. Only lipid species were used, which had at least 2 valid measurements in each of the two cell lines (n = 1064). The FADS3 KO clone C02 is displayed, which was created by the deletion method.