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. 2023 Dec 4;25(12):1821–1832. doi: 10.1038/s41556-023-01274-x

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© The Author(s) 2023

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 — a, [U-¹³C]glucose tracer analysis of vehicle- and 10 μM UK5099-treated basal-derived organoids 7 d after plating (n = 3 independent biological replicates). Data are shown as mean ± s.e.m. b,c, Percentage organoid formation (n = 3 independent biological replicates) (b) and organoid diameter (n = 25 independent biological samples) (c) of vehicle- and 10 μM UK5099-treated basal-derived organoids 7 d after plating. d, Western blot analysis of luminal markers androgen receptor (AR) and KRT8 and basal marker p63 in vehicle- and 10 μM UK5099-treated basal-derived organoids 7 d after plating. e, Immunofluorescence of luminal marker KRT8 and basal marker p63 in representative vehicle- and 10 μM UK5099-treated basal-derived organoids 7 d after plating. Scale bars, 100 μm. f, Intracellular flow cytometry of KRT8 and basal marker cytokeratin 5 (KRT5) in vehicle- and 10 μM UK5099-treated basal-derived organoids 7 d after plating. g, Quantification of mean fluorescence intensity (MFI) of KRT8 from panel f (n = 4 independent biological replicates). h, [U-¹³C]glucose tracer analysis of control and Mpc1-KO basal-derived organoids (n = 3 independent biological replicates). Data are shown as mean ± s.e.m. i, Western blot analysis of basal and luminal markers in control and Mpc1-KO basal-derived organoids. j, GSEA showing negative enrichment of CD49f^low luminal signature¹⁸ in Mpc1-KO relative to control basal-derived organoids. k, Flow cytometry quantification of percentage of EpCAM⁻KRT8⁻ cells in vehicle- and 10 μM UK5099-treated primary and quaternary basal-derived organoids (n = 3 independent biological replicates). l, t-Distributed stochastic neighbour embedding (t-SNE) plot of scRNA-seq data on quaternary prostate organoids illustrating distinct cell populations. m, t-SNE plot of vehicle- and 10 μM UK5099-treated cells from scRNA-seq data. n, Quantification of percentage of vehicle- and 10 μM UK5099-treated cells in each cluster from scRNA-seq data. For all panels, error bars represent s.e.m. P values were calculated using an unpaired two-tailed t-test with Welch’s correction. Aco, aconitate; Cit, citrate; EMT, epithelial–mesenchymal transition; Lac, lactate; Mal, malate; orgs, organoids; Succ, succinate; UK, UK5099; Veh, vehicle.

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