choline deficient mcd diet  (Valiant Co Ltd)

Journal: bioRxiv

Article Title: Flexibility of systemic one-carbon metabolism partially buffers dietary methyl donor deficiency

doi: 10.64898/2026.02.05.703880

Figure Lengend Snippet: (a) Schematic showing intravenous infusion of [trimethyl- H 9 ]choline and [methyl- C 1 ]methionine in mice fed a methionine and choline deficient diet or fed a paired control diet. (b) Rate of appearance of choline calculated from m+9 serum enrichments from 8 h intravenous infusion of [trimethyl- H 9 ]choline. (c) Rate of appearance of methionine calculated from m+1 serum enrichments from 8 h intravenous infusion of [methyl- C 1 ]methionine. (d) Normalized labeling of tissue SAM from 8 hour intravenous infusion of [methyl- C 1 ]methionine. (e) Normalized labeling of tissue SAM from 8 hour intravenous infusion of [trimethyl- H 9 ]choline. The red line represents the fraction of serum methionine that is labeled in control conditions and the green line represents the fraction of serum methionine that is labeled in MCD diet. Labeling below these lines indicates that labeling is potentially sourced from serum methionine pools rather than tissue autonomous production. (f) Normalized labeling of pooled PCs in the liver from 8 hour intravenous infusion of [trimethyl- H 9 ]choline. (g) Ratio of ion counts of pooled PC species to pooled PE species. Bars represent mean ± S.D. Each data point represents data from an independent mouse. P-values were calculated with two-way ANOVA with Šídák correction for multiple comparisons. All experiments were performed in male ad lib fed C57Bl/6N mice during the light cycle. PC, phosphatidylcholine; PE, phosphatidylethanoline; PEMT, phosphatidylethanolamine methyltransferase

Article Snippet: Methyl deficiency was induced by transitioning mice from standard chow to a methionine and choline-deficient (MCD) diet (MP Biomedicals, MP296043910), while age-matched controls received a matched methionine and choline replete control diet (MP Biomedicals, MP296044110).

Techniques: Control, Labeling

Journal: bioRxiv

Article Title: Flexibility of systemic one-carbon metabolism partially buffers dietary methyl donor deficiency

doi: 10.64898/2026.02.05.703880

Figure Lengend Snippet: (a) Normalized labeling of serum and tissue serine from 8 hour intravenous infusion of [trimethyl- H 9 ]choline.The red line represents the fraction of serum serine that is labeled in control conditions and the green line represents the fraction of serum serine that is labeled in MCD diet. Labeling below these lines indicates that labeling is potentially sourced from serum serine pools rather than tissue autonomous production. (b) Relative ion counts for glycine across tissues in MCD diet and control diet fed mice. Bars represent mean ± S.D. Each data point represents data from an independent mouse. P-values were calculated with two-way ANOVA with Šídák correction for multiple comparisons (a) or Mixed-effects analysis with Šídák correction for multiple comparisons (b). All experiments were performed in male ad lib fed C57Bl/6N mice during the light cycle. MCD, methionine and choline deficient

Article Snippet: Methyl deficiency was induced by transitioning mice from standard chow to a methionine and choline-deficient (MCD) diet (MP Biomedicals, MP296043910), while age-matched controls received a matched methionine and choline replete control diet (MP Biomedicals, MP296044110).

Techniques: Labeling, Control

Journal: bioRxiv

Article Title: Flexibility of systemic one-carbon metabolism partially buffers dietary methyl donor deficiency

doi: 10.64898/2026.02.05.703880

Figure Lengend Snippet: (a) Schematic of the folate cycle highlighting reactions that support production of 5,10-methylene-THF and SAM. (b) Schematic showing intravenous infusion of [2,3,3- H 3 ]serine and [U- C ]glycine in mice fed a methionine and choline deficient diet or fed a paired control diet. (c) Rate of appearance of serine calculated from m+3 serum enrichments from 8 h intravenous infusion of [2,3,3- H 3 ]serine. (d) Rate of appearance of glycine calculated from m+2 serum enrichments from 8 h intravenous infusion of [U- C 2 ]glycine. (e) Schematic of use of [2,3,3- H 3 ]serine tracing to measure the contribution of the folate cycle to SAM synthesis. (f) Normalized labeling of tissue SAM from 8 hour intravenous infusion of [2,3,3- H 3 ]serine. The red line represents the fraction of serum methionine that is labeled in control conditions and the green line represents the fraction of serum methionine that is labeled in MCD diet. Labeling below these lines indicates that labeling is potentially sourced from serum methionine pools rather than tissue autonomous production. Bars represent mean ± S.D. Each data point represents data from an independent mouse. P-values were calculated with two tailed T-test (c,d) or two-way ANOVA with Šídák correction for multiple comparisons (f). All experiments were performed in male ad lib fed C57Bl/6N mice during the light cycle. MCD, methionine and choline deficient; SHMT, serine hydroxymethyltransferase, MTR, 5-methyltetrahyrofolate homocysteine methyltransferase; SAM, s-adenosylmethionine

Article Snippet: Methyl deficiency was induced by transitioning mice from standard chow to a methionine and choline-deficient (MCD) diet (MP Biomedicals, MP296043910), while age-matched controls received a matched methionine and choline replete control diet (MP Biomedicals, MP296044110).

Techniques: Control, Labeling, Two Tailed Test

Journal: bioRxiv

Article Title: Flexibility of systemic one-carbon metabolism partially buffers dietary methyl donor deficiency

doi: 10.64898/2026.02.05.703880

Figure Lengend Snippet: (a) Schematic showing potential sources of circulating serine. (b) Normalized labeled fraction of serum serine from infusion of [U- C 2 ]glycine. (c) Normalized labeled fraction of serum serine from infusion of [U- C 6 ]glucose. (d) Rate of appearance of essential amino acid phenylalanine from intravenous infusion of [ N]phenylalanine. (e) Serine production fluxes from different sources measured using the serine rate of appearance and infusions of U- C 2 ]glycine, U- C 6 ]glucose, and [ N]phenylalanine. (f) Relative ion counts for serine across tissues in MCD diet and control diet fed mice. (g) Schematic of proposed mechanism. When methionine and choline are deficient in the diet, there is increased release of serine from the kidney that supports maintained methionine, PC, and choline synthesis in the liver. Thus, methionine and choline fluxes are maintained by one-carbon metabolism buffering. Bars represent mean ± S.D. Each data point represents data from an independent mouse. P-values were calculated with two tailed T-test (b), two-way ANOVA with Šídák correction for multiple comparisons (e), or mixed-effects analysis with Šídák correction for multiple comparisons (f). All experiments were performed in male ad lib fed C57Bl/6N mice during the light cycle. SAM, s-adenosylmethionine; MCD, methionine and choline deficient

Article Snippet: Methyl deficiency was induced by transitioning mice from standard chow to a methionine and choline-deficient (MCD) diet (MP Biomedicals, MP296043910), while age-matched controls received a matched methionine and choline replete control diet (MP Biomedicals, MP296044110).

Techniques: Labeling, Control, Two Tailed Test

Journal: JHEP Reports

Article Title: Hepatic immune environment differences among common mouse strains in models of MASH and liver cancer

doi: 10.1016/j.jhepr.2025.101380

Figure Lengend Snippet: Changes of liver immune subsets under MASH in BALB/c, C57BL/6, and FVB/N mice fed with MCD diet. Female BALB/c, C57BL/6, and FVB/N mice were fed with MCD diet ( vs. regular diet) to induce MASH. (A) The development of MASH was confirmed by H&E staining. Liver immune cells from MASH mice or control mice were prepared and immune subsets were measured by flow cytometry analysis. The comparison between MCD (green) and control (gray) was performed in each liver immune subsets of the three mouse strains (B–G). The overall changes of various liver immune subsets from three mouse strains are shown (H). The size of circle represents the log 10 transformed fold changes of each immune subset. The color gradient represents the relative frequences of each immune subset. The distribution of fold changes of each type of immune cell is also shown; n = 4 per group, two-way ANOVA with Bonferroni correction, ∗ p <0.05. MASH, metabolic dysfunction-associated steatohepatitis; MCD diet, methionine- and choline-deficient diet; Tregs, regulatory T cells.

Article Snippet: MASH was induced in mice by feeding a methionine- and choline-deficient (MCD) diet (Research diets inc., New Brunswick, NJ, USA Ref. A02082002BR) for a period of 3 weeks, or a Western diet (Envigo, Indianapolis, IN, USA, Ref. TD.120528) with high sugar solution (23.1 g/L d-fructose and 18.9 g/L d-glucose) with weekly intraperitoneal injections of carbon tetrachloride (CCl 4 ) (Sigma, St. Louis, MO, USA, Cat# 289116) at a dose of 0.32 mg/g of body weight for 12 weeks as previously reported.

Techniques: Staining, Control, Flow Cytometry, Comparison, Transformation Assay

Journal: JHEP Reports

Article Title: Hepatic immune environment differences among common mouse strains in models of MASH and liver cancer

doi: 10.1016/j.jhepr.2025.101380

Figure Lengend Snippet: Cross-species comparison of liver immune changes by MASH between mice and humans. The published human scRNA-seq dataset GSE159977 of CD45+ cells from either MASH or healthy livers were processed using Seurat (5.1.0). (A) shows the UMAP split based on healthy or MASH. (B) shows the dot plot of marker genes for each annotated cell clusters. (C) Liver CD45 + cell compositions were measured in naïve BALB/c, C57BL/6, and FVB/N strains by flow cytometry as described in Fig. 1 A. CD45 + cell composition of healthy human liver or human HCC adjacent liver tissues were calculated based on the scRNA-seq datasets of GSE159977 or phs003279.v1.p1, respectively. (D,E) CD45 + or CD4 + T cell compositions of MASH or healthy human livers were calculated from GSE159977 . (F) In total liver CD45 + cells, the frequencies of shared major liver immune subsets between mice and human, including CD8+ T cell, CD4+ T cell, B cells, γδT cells and NK cells, were calculated in MASH or healthy human livers ( GSE159977 ) and MASH or control mice of three strains under either MCD diet or Western + CCl 4 diet MASH model. CCl 4 , carbon tetrachloride; HCC, hepatocellular carcinoma; MASH, metabolic dysfunction-associated steatohepatitis; MAIT cells, mucosal-associated invariant T cells; MCD diet, methionine- and choline-deficient diet; NK, natural killer; UMAP, Uniform Manifold Approximation and Projection.

Article Snippet: MASH was induced in mice by feeding a methionine- and choline-deficient (MCD) diet (Research diets inc., New Brunswick, NJ, USA Ref. A02082002BR) for a period of 3 weeks, or a Western diet (Envigo, Indianapolis, IN, USA, Ref. TD.120528) with high sugar solution (23.1 g/L d-fructose and 18.9 g/L d-glucose) with weekly intraperitoneal injections of carbon tetrachloride (CCl 4 ) (Sigma, St. Louis, MO, USA, Cat# 289116) at a dose of 0.32 mg/g of body weight for 12 weeks as previously reported.

Techniques: Comparison, Marker, Flow Cytometry, Control, Western Blot