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. 2018 Nov 16;1:194. doi: 10.1038/s42003-018-0200-x

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© The Author(s) 2018

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. 5 — Glutaminolysis and fatty acids synthase dependence of Pkd1^−/− cells. a The scheme illustrates the fate of glutamine C atoms in TCA cycle intermediates and fatty acid biosynthesis. b Quantification of intracellular citrate (M + 5) labelled by glutamine was significantly higher in Pkd1^−/− as compared with Pkd1^+/+ cells. c Incorporation of glutamine carbons into intracellular palmitate (M + 2) was significantly higher in Pkd1^−/− as compared with Pkd1^+/+ cells. Data are means from six technical replicates from one experiment. d Dot plots showing means fold change, SEM, mRNA levels of Fasn in Pkd1^−/− compared to mock Pkd1^+/+ cells and SiFasnPkd1^−/− compared to mock Pkd1^−/⁻ normalized to Hprt. Three technical replicates of at least 3 independent experiments. e Dot plots showing means fold change, SEM, mRNA Fasn levels measured in control compared to cystic kidneys normalized to Hprt. n = 8, two technical replicates of at least two independent experiments. f Representative images showing cellular morphology of siFasn in Pkd1^+/+ and Pkd1^−/− and mock controls. g Representative graph of percentage Ki67-positive cells for mock and SiFasn for Pkd1^+/+ and Pkd1^−/− cells after 48 h of transient silencing. Data from three technical replicates of at least two independent experiments. h Representative graphs showing percentage of apoptotic cells after 48 h of transfection, data from three technical replicates from three independent experiments. i Lipidomic profiling of cystic versus control kidneys showed altered percentage of diacylglycerol (DAG:38.4), three different species of triacylglycerol (TAG:50, TAG:52 TAG:54) and sterol esters (SE:43) from total lipid content, n = 6. Scale bar is 100 μm. Mean ± SEM were indicated, n.s. not significant (p ≥ 0.05), *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. t-test for b, c, ANOVA followed by Bonferroni for d, e, g, and h ANOVA for i