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. 2021 Jan 14;12:366. doi: 10.1038/s41467-020-20223-y

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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 Percentage of BrdU positive cells in HCT116 and DLD-1 cells grown in CM or -SG medium ±10 µM PH755 for 48 h followed by a 5-h incubation with 10 µM BrdU. Data represents mean ± SEM of three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA with Tukey’s post hoc test). b Percentage of SubG1 cells in HCT116 and DLD-1 cells grown in CM or -SG medium ±10 µM PH755 for 48 h. Data represents mean ± SEM of five independent experiments (**p < 0.01, ***p < 0.001, one-way ANOVA with Tukey’s post hoc test). c Cells were grown in CM or -SG medium supplemented or not with 10 µM PH755 for 2 days (HCT116) or 3 days (DLD-1). Western blots show the expression of cleaved Caspase-3 and Caspase-3. Membrane was reprobed with vinculin as a loading control. Data are representative of three independent experiments. d Intracellular serine level in cells grown in CM or -SG medium ±10 µM PH755 containing U-[¹³C]-glucose was measured by LC-MS. Metabolite percentages are represented as mean ± SEM of triplicate cultures and are representative of three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; one-way ANOVA with Tukey’s post hoc test). e Intestinal tumour organoids derived from Vil1-creER;Apc^fl/fl (Apc) and Vil1-creER;Apc^fl/fl;Kras^G12D/+ (Apc Kras) mice were grown in CM or -SG medium supplemented or not with 10 µM PH755. Left panel: representative pictures of the organoids are shown before (day 0) and 2 days after medium change. Right panel: quantification of organoid diameter 2 (Apc) or 4 days (Apc Kras) after medium change. Data are presented as mean ± SEM (n = number of organoids measured per condition; Apc: CM: n = 113, CM + PH755: n = 200, -SG: n = 190, -SG + PH755: n = 158; Apc Kras: CM: n = 149, CM + PH755: n = 134, -SG: n = 134, -SG + PH755: n = 78) and are representative of at least two independent experiments (***p < 0.001, ****p < 0.0001; one-way ANOVA with Tukey’s post hoc test). Scale bar represents 200 μm.