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. 2024 Jan 2;15:79. doi: 10.1038/s41467-023-44412-7

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© The Author(s) 2024

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. 2 — a Volcano plot displays log2 fold-change (FC) in lipid species abundance, comparing nocodazole-treated to vehicle-treated Caki-1 cells (n = 510, P < 0.05, FC > 1.5). AcCa, acyl carnitine; CE, cholesterol ester; CER, ceramide; CL, cardiolipin; Co, coenzyme; DAG, diacylglyceride; HexCer, hexosyl ceramides; LPC, lysophosphatidylcholine; LPE, lysophosphatidylethanolamine; LPG, lysophosphatidylglycerol; LPI, lysophosphatidylinositol; PC, phosphatidylcholine; PC-O, ether-linked phosphatidylcholine; PE, phosphatidylethanolamine; PE-O/PE-P, ether-linked phosphatidylethanolamine; PA, phosphatidic acid; PG, phosphatidylglycerol; PI, phosphatidylinositol; PS, phosphatidylserine; SM, sphingomyelin; TAG, triacylglyceride; TAG-O, ether-linked triacylglyceride. b Boxplots displaying log2 FC in non-TAG (n = 413) and TAG (n = 97) lipid species abundance between nocodazole-treated and vehicle-treated Caki-1 cells, with medians and whiskers indicating min-max values. BODIPY 493/503 staining in Caki-1 cells after 48 h of cell cycle inhibitor treatment. scale bar, 20 μm (c), and quantifications in the indicated cell lines (d). BODIPY 493/503 staining in Caki-1 cells treated with iCDK4/6 for 48 h (e), or WT and sgCDK1 cells (f). g Triglyceride concentrations in Caki-1 cells. h Heat map of the significantly changed lipids (unpaired, two-tailed t-test; FDR-corrected P < 0.05, fold change > 1.5 in vehicle- versus nocodazole-treated Caki-1 cells). Rows represent samples, columns represent normalized intensities, color-coded from red (high intensity) to blue (low intensity). Volcano plots for iDGAT1/2-treated versus vehicle-treated (i, n = 276) and iDGAT1/2 and nocodazole-treated versus nocodazole-treated (j, n = 302) Caki-1 cells. Log2 FC in phospholipid abundance comparing iDGAT and nocodazole-treated to nocodazole-treated Caki-1 cells, depicted with boxplots (k) and volcano plots (l). n = 192. LPC-O, ether-linked lysophosphatidylcholine; LPS, lysophosphatidylserine. BODIPY 493/503 staining in Caki-1 (m) and ACHN (n) cells. o Immunoblot of DGAT1 in Caki-1 cells. p BODIPY 493/503 staining in WT, DGAT DKO1, and DKO2 Caki-1 cells with or without exposure to hydroxyurea (0.3 mM) for 48 h. Mean ( ± SD) values are shown. n = 3. n indicates independent repeats, except a, b, i, j, k, and l. unpaired, two-tailed t-test. n.s. not significant. Source data are provided as a Source Data file.