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. 2023 Dec 8;5(12):2148–2168. doi: 10.1038/s42255-023-00936-2

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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. 7 — a, Schematic of FACS sorting strategy from DOX-inducible iCas9-GFP; PHGDH-depleted HCT116 cells expressing either a combination of NTCs, SLC6A14/12A4 or SLC6A14/25A15 gRNAs. b, Growth curves of cells from a. Data are mean ± s.d. from n = 3 biological replicates per condition for each time point. c, Schematic of in vivo experiments. d, Tumor volume over time of tumors from PHGDH-depleted HCT116 cells expressing NTC gRNAs ±DOX. Values are mean ± s.e.m. from n = 9 −DOX and n = 11 +DOX mice. e, Kaplan–Meier plot showing probability of survival on mice from d. f, Tumor volume over time of tumors from PHGDH-depleted HCT116 cells expressing SLC6A14/12A4 gRNAs ±DOX. Values are mean ± s.e.m. from n = 11 −DOX and n = 12 +DOX treated mice. g, Kaplan–Meier plot showing probability of survival on mice from f. h, Serine levels in tumors formed by PHGDH-depleted HCT116 cells expressing NTC, SLC6A14/SLC12A4 or SLC6A14/SLC25A15 gRNAs with (left) or without (right) DOX induction. Values are mean ± s.d. from n = 6 mice per condition. i, Serine levels in circulating blood serum of mice with tumors formed by PHGDH-depleted HCT116 cells expressing NTC, SLC6A14/SLC12A4 or SLC6A14/SLC25A15 gRNAs with (left) or without (right) DOX induction. Values are mean ± s.d. from n = 6 mice per condition. j, Glycine levels in tumors formed by PHGDH-depleted HCT116 cells expressing NTC, SLC6A14/SLC12A4 or SLC6A14/SLC25A15 gRNAs with (left) or without (right) DOX induction. Values are mean ± s.d. from n = 6 mice per condition. k, Glycine levels in circulating blood serum of mice with tumors formed by PHGDH-depleted HCT116 cells expressing NTC, SLC6A14/SLC12A4 or SLC6A14/SLC25A15 gRNAs with (left) or without (right) DOX induction. Values are mean ± s.d. from n = 6 mice per condition. l, Schematic summary. Statistical significance was assessed with two-tailed unpaired t-test (d,f), two-tailed Gehan–Breslow–Wilcoxon test (e,g) and ordinary ANOVA and Dunnett’s multiple comparisons test (h–k).