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. 2021 Oct 28;11:21268. doi: 10.1038/s41598-021-99958-7

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Flcn loss reduces MCD diet-induced fibrosis and inflammation. (A) Relative quantitative real‐time PCR analysis of fibrosis-related genes mRNA transcript levels in livers of mice fed either on chow or methionine/choline deficient diet (MCD) over a period of 6 weeks (mean ± SEM of the RNA fold change of indicated mRNAs; two-way ANOVA, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; n = 8 mice per condition). (B) Sirius red staining in liver sections of mice fed as described in (A). Images are representative of 6 mice per condition. (C) Quantification of the sirius staining in liver sections of mice fed as described in (A) (mean ± SEM, two-way ANOVA, **P < 0.01; ****P < 0.0001; n = 6 mice per condition). (D) α-SMA Immunohistochemistry (IHC) staining in liver sections of mice fed as described in (A). Data are representative of 4 mice per condition. (E) Quantification of α-SMA IHC staining in liver sections of mice fed as described in (A) (mean ± SEM, two-way ANOVA, **P < 0.01; ***P < 0.001; n = 4 mice per condition). (F) 4-HNE Immunohistochemistry (IHC) staining in liver sections of mice fed as described in (A). Data are representative of 4 mice per condition. (G) Quantification of 4-HNE IHC staining in liver sections of mice fed as described in (A) (mean ± SEM, two-way ANOVA, **P < 0.01; ***P < 0.001, ****p < 0.0001; n = 4 mice per condition). (H) Relative quantitative real‐time PCR analysis of inflammation-related genes mRNA transcript levels in livers of mice fed as described in (A) (mean ± SEM of the RNA fold change of indicated mRNAs; two-way ANOVA, *P < 0.05, **P < 0.01, ****P < 0.0001; n = 8 mice per condition). (I-J) Mouse cytokines and chemokines quantification in liver homogenates of mice fed as described in (A) (mean ± SEM, two-way ANOVA, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; n = 3 mice per condition). (K) F4/80 Immunohistochemistry (IHC) staining in liver sections of mice fed as described in (A). Data are representative of 4 mice per condition. (L) Quantification of F4/80 IHC staining in liver sections of mice fed as described in (A) (mean ± SEM, two-way ANOVA, *P < 0.05; **P < 0.01; n = 4 mice per condition).