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. 2021 Dec 2;12:7033. doi: 10.1038/s41467-021-26840-5

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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. 2 — a Projection of the indicated human cell lines ex vivo and patient-derived xenograft (PDX) models in vivo to the pseudotime (PT) inferred trajectory. Normal prostate: green; hormone-sensitive tumors (HSPC): red; castration-resistant (CRPC) tumors: blue; neuroendocrine (NEPC) tumors: violet; ex vivo cell lines: yellow. Representative replicates for each sample are shown. See Source data file. b Immunoblotting analysis of the indicated proteins across PDX models indicates an upregulation of polycomb-repressive complex-2 members and G2M cell cycle checkpoint genes. Experiments were repeated at least three times with similar results. See Source data file. c Androgen-dependent xenograft models progress along the trajectory when developing castration resistance. Normal prostate: green; hormone-sensitive tumors (HSPC): red; castration-resistant (CRPC) tumors: blue; neuroendocrine (NEPC) tumors: violet; pre-castration (Pre-CX) PDX models: magenta; post castration (Post-CX) PDX models: dark blue; Wk weeks; s7, s9, and s10 indicate different clones derived from LuCaP-23.1 PDX model. See Source data file. d Immunoblot analysis of LNCaP xenograft models shows upregulation of AR, polycomb-repressive complex-2 members, and G2M cell cycle checkpoint genes upon recurrence after castration (Cx). Experiments were repeated at least three times with similar results. See Source data file. e Three-dimensional (3D) ex vivo cultures in Matrigel of the indicated PDX and the xenografted LNCaP cells show higher PT than their in vivo counterparts. Normal prostate: green; hormone-sensitive tumors (HSPC): red; castration-resistant (CRPC) tumors: blue; neuroendocrine (NEPC) tumors: violet; ex vivo: yellow. Representative replicates for each sample are shown. Individual replicates are depicted in Supplementary Fig. 2. See Source data file. f Dihydrotestosterone (DHT) dose–response curves of the indicated models in 3D using Matrigel versus standard 2D culture. In the 3D conditions, DHT dependency is largely abolished. For each curve, n = 3 biologically independent experiments are reported (three animals for each curve). Error bars represent standard errors for each time point. LNCaP: red; PNPCa, LuCaP and PNPCa models: gray. See Source data file.