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. 2023 May 19;8:187. doi: 10.1038/s41392-023-01405-8

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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. 6 — CSN6/FASN-induced lipogenesis promotes CRC tumor growth, which can be inhibited by orlistat. a The effect of orlistat on CSN6-mediated lipogenesis was determined by BODIPY 493/503 staining followed by flow cytometry analysis. Representative histograms and mean fluorescence intensities (MFI) in HCT 116 (left) and DLD-1 (right) cells were shown. b Cell growth assays for HCT 116 (left) and DLD-1 (right) cells stably transfected with indicated plasmids and then treated with vehicle (DMSO) or orlistat (8 μM). c Western blot analysis for cleaved PARP1 expression in HCT 116 and DLD-1 cells stably transfected with indicated plasmids and then treated with vehicle (DMSO) or orlistat (8 μM). Arrow denotes molecular weight of cleaved PARP1. d, e Colony-formation assays (d) and migration assays (e) generated from HCT 116 and DLD-1 cells stably transfected with indicated plasmids and then treated with vehicle (DMSO) or orlistat (8 μM). Scale bar, 200 μm. f Scheme of experimental design for DLD-1 subcutaneous tumor model (top). Tumor growth curves of indicated DLD-1 tumors (n = 5 mice per group) were shown (bottom). g Bioluminescence overlay images of indicated DLD-1 tumors in mice after 28 days growth (left). Colored scale shows photon flux in units of radiance (photons/s/cm²/steradian(sr)). Quantification of photon flux for the indicated DLD-1 tumors was shown (right). h The resulting tumors generated in f were resected and photographed at the end of the experiment (top). Tumor weights of indicated DLD-1 tumors were shown (bottom). i Representative IHC images of CSN6, FASN and Ki67 staining in indicated DLD-1 tumors (left). The regions in black boxes are shown at higher magnification. Scale bars, 50 μm. Quantification of the percentage of CSN6 positive, FASN positive and Ki67 positive cells of indicated groups was shown (right). j Fluorescence microscopy analysis of frozen sections from indicated DLD-1 tumors stained with BODIPY 493/503 dye (green) and DAPI (blue). Representative images (left) and fluorescent BODIPY quantification (right) of indicated groups were shown. Scale bar, 20 μm. k Schematic design for in vivo fatty acid metabolite tracing of indicated DLD-1 tumors (top). Ratio of ¹³C-Glucose to indicated fatty acid metabolites was shown (bottom). All values are expressed as means ± SD. ns not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; as determined by two-way ANOVA (b, f), by one-way ANOVA (a, d, e, g–j) or by two-sided Student’s t-test (k)