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Addgene inc human mat2a
Summary of V/K Kinetic Isotope Effects, Intrinsic Kinetic Isotope Effects, Binding Isotope Effects and Calculated Kinetic Isotope Effects from QM Calculations for <t> Human MAT2A </t> at 5 Atomic Positions

Summary of V/K Kinetic Isotope Effects, Intrinsic Kinetic Isotope Effects, Binding Isotope Effects and Calculated Kinetic Isotope Effects from QM Calculations for <t> Human MAT2A </t> at 5 Atomic Positions

Human Mat2a, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human mat2a/product/Addgene inc
Average 92 stars, based on 4 article reviews

human mat2a - by Bioz Stars, 2026-04

92/100 stars

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1) Product Images from "The Transition-State Structure for Human MAT2A from Isotope Effects"

Article Title: The Transition-State Structure for Human MAT2A from Isotope Effects

Journal: Journal of the American Chemical Society

doi: 10.1021/jacs.7b05803

Figure Legend Snippet: Summary of V/K Kinetic Isotope Effects, Intrinsic Kinetic Isotope Effects, Binding Isotope Effects and Calculated Kinetic Isotope Effects from QM Calculations for Human MAT2A at 5 Atomic Positions

Techniques Used: Binding Assay

MAT2A catalyzes the formation of SAM from ATP and methionine. KIE measurements used isotopically labeled substrates with the labels indicated. Each color corresponds to labels found on an individual substrate molecule.

Figure Legend Snippet: MAT2A catalyzes the formation of SAM from ATP and methionine. KIE measurements used isotopically labeled substrates with the labels indicated. Each color corresponds to labels found on an individual substrate molecule.

Techniques Used: Labeling

(A) Geometric and electrostatic potential surface (EPS) maps for the MAT2A transition state. Red color designates partial negative charge while blue color designates partial positive charge. (B) Zoomed in EPS maps of the central sulfur atom for the MAT2A transition state (TS), S-adenosyl-L-methionine (SAM) and methionine (Met) are shown for comparison. The partial positive charge on the sulfonium group is significantly reduced in the transition state structure when compared to the SAM structure.

Figure Legend Snippet: (A) Geometric and electrostatic potential surface (EPS) maps for the MAT2A transition state. Red color designates partial negative charge while blue color designates partial positive charge. (B) Zoomed in EPS maps of the central sulfur atom for the MAT2A transition state (TS), S-adenosyl-L-methionine (SAM) and methionine (Met) are shown for comparison. The partial positive charge on the sulfonium group is significantly reduced in the transition state structure when compared to the SAM structure.

Techniques Used: Comparison

Image Search Results

A Score plot depicting the separation of metabolic gene patterns in the human DW and NDW groups through PCA analysis in GSE154556 . B Volcano plot showing the differentially expressed metabolic genes between human DWs and NDWs in GSE154556 . Differentially expressed genes were assessed with the limma moderated two-sided t test. C KEGG analysis of typical differential metabolic pathways between human DWs and NDWs in GSE154556 . D t-SNE plots of the characterized cell clusters identified via scRNA-seq of human wound samples ( GSE165816 ). E Venn diagram showing the shared altered metabolic differential genes and their origins. F Correlation analysis of the expression levels of the metabolic candidates and the inflammatory macrophage infiltration score in GSE154556 . The text annotations above showed the cellular origins of the main differences of these candidates analyzed from GSE165816 . G Cellular communication analysis revealing potential interactions among pericytes with low MAT2A expression and other cell types from GSE165816 . H Schematic illustration of the methionine cycle, and the relative levels of methionine in the human DW and NDW groups. n = 12 biologically independent samples. I Expression levels of metabolic enzymes involved in the methionine cycle in the two groups ( GSE165816 ). Non-parametric two-sided Wilcoxon rank-sum test was used. J Immunofluorescence staining and statistical analysis demonstrating the expression levels of MAT2A in CD31-NG2+PDGFRβ+ pericytes from human wounds. n = 3 biologically independent samples. K Pericytes were classified into samples with high MAT2A expression levels and samples with low MAT2A expression levels ( GSE165816 ); grouped samples were analyzed via GSEA. The median expression of the gene was used as the dividing line. Data were shown as mean ± SD. Statistical significance was determined using hypergeometric test ( C ) and two-tailed unpaired t test ( H , J ). Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: A biomimetic senotherapy replenishing MAT2A promotes wound regeneration in preclinical models

doi: 10.1038/s41467-025-65659-2

Figure Lengend Snippet: A Score plot depicting the separation of metabolic gene patterns in the human DW and NDW groups through PCA analysis in GSE154556 . B Volcano plot showing the differentially expressed metabolic genes between human DWs and NDWs in GSE154556 . Differentially expressed genes were assessed with the limma moderated two-sided t test. C KEGG analysis of typical differential metabolic pathways between human DWs and NDWs in GSE154556 . D t-SNE plots of the characterized cell clusters identified via scRNA-seq of human wound samples ( GSE165816 ). E Venn diagram showing the shared altered metabolic differential genes and their origins. F Correlation analysis of the expression levels of the metabolic candidates and the inflammatory macrophage infiltration score in GSE154556 . The text annotations above showed the cellular origins of the main differences of these candidates analyzed from GSE165816 . G Cellular communication analysis revealing potential interactions among pericytes with low MAT2A expression and other cell types from GSE165816 . H Schematic illustration of the methionine cycle, and the relative levels of methionine in the human DW and NDW groups. n = 12 biologically independent samples. I Expression levels of metabolic enzymes involved in the methionine cycle in the two groups ( GSE165816 ). Non-parametric two-sided Wilcoxon rank-sum test was used. J Immunofluorescence staining and statistical analysis demonstrating the expression levels of MAT2A in CD31-NG2+PDGFRβ+ pericytes from human wounds. n = 3 biologically independent samples. K Pericytes were classified into samples with high MAT2A expression levels and samples with low MAT2A expression levels ( GSE165816 ); grouped samples were analyzed via GSEA. The median expression of the gene was used as the dividing line. Data were shown as mean ± SD. Statistical significance was determined using hypergeometric test ( C ) and two-tailed unpaired t test ( H , J ). Source data are provided as a Source Data file.

Article Snippet: D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration.

Techniques: Expressing, Immunofluorescence, Staining, Two Tailed Test

A ScRNA-seq profiling workflow created with BioRender.com. B Western blot analysis of MAT2A expression in pericytes isolated from the indicated mouse skin. C Representative images of cutaneous wounds of mice on days 0, 4, 8, 12, and 16 after wound model generation by surgical excision. Ratio of wound sizes were quantified by using ImageJ software and were calculated by the percentages of wound closure compared to day 0 wound size. n = 3 mice for sampling at the indicated time points. D Representative blood perfusion images and statistical analysis of wounds at days 4 and 8 after surgery. E Cutaneous wound sections were subjected to H&E and Masson’s trichrome staining, and IHC staining for Ki-67, α-SMA, and IL6 were performed. Samples were collected at day 8 after wound model generation. n = 3 mice for sampling at the indicated time points. Scale bar, 100 μm. F UMAP plot showing identified cell clusters of mouse skin. G Representative immunofluorescence images and statistical analysis demonstrating the pericyte abundance in cutaneous wounds on day 4. H Volcano plot showing the differential genes of pericyte clusters between the two groups based on P value < 0.05 and absolute log 2 (Fold Change) > 0.25. Non-parametric two-sided Wilcoxon rank-sum test was used. I Bar graph showing the functionally enriched pathways associated with the significantly upregulated or downregulated genes (MAT2A LOF vs. control) in the pericyte clusters. J Cellular communication analysis revealing potential interactions among pericytes and other cell types. K Violin-box plots representing the expression of proinflammatory or anti-inflammatory signature genes in the macrophage clusters. Macrophage cells in control group, n = 6201; macrophage cells in MAT2A LOF group, n = 6763. L UMAP plot and quantitative analysis showing characterized cell clusters of infiltrated macrophages and their proportions in the indicated groups. M Cell trajectory analysis of the characterized cell clusters of infiltrated macrophages. N Representative immunofluorescence images and statistical analysis demonstrating the macrophageinfiltration in cutaneous wound tissues on day 8. n = 3 mice in each group. Scale bar, 20 μm. For the box and violin-box plots in ( G ), and ( K ), the centerlines indicated the medians. The box limits indicated the first and third quartiles. The whiskers indicated the maxima and minima. Data in the bar plots were shown as mean ± SD. n = 3 biologically independent samples ( D , G ). Statistical significance was determined using two-way ANOVA with Tukey’s multiple comparisons test ( C ) and two-tailed unpaired t test ( D , G , K , N ). Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: A biomimetic senotherapy replenishing MAT2A promotes wound regeneration in preclinical models

doi: 10.1038/s41467-025-65659-2

Figure Lengend Snippet: A ScRNA-seq profiling workflow created with BioRender.com. B Western blot analysis of MAT2A expression in pericytes isolated from the indicated mouse skin. C Representative images of cutaneous wounds of mice on days 0, 4, 8, 12, and 16 after wound model generation by surgical excision. Ratio of wound sizes were quantified by using ImageJ software and were calculated by the percentages of wound closure compared to day 0 wound size. n = 3 mice for sampling at the indicated time points. D Representative blood perfusion images and statistical analysis of wounds at days 4 and 8 after surgery. E Cutaneous wound sections were subjected to H&E and Masson’s trichrome staining, and IHC staining for Ki-67, α-SMA, and IL6 were performed. Samples were collected at day 8 after wound model generation. n = 3 mice for sampling at the indicated time points. Scale bar, 100 μm. F UMAP plot showing identified cell clusters of mouse skin. G Representative immunofluorescence images and statistical analysis demonstrating the pericyte abundance in cutaneous wounds on day 4. H Volcano plot showing the differential genes of pericyte clusters between the two groups based on P value < 0.05 and absolute log 2 (Fold Change) > 0.25. Non-parametric two-sided Wilcoxon rank-sum test was used. I Bar graph showing the functionally enriched pathways associated with the significantly upregulated or downregulated genes (MAT2A LOF vs. control) in the pericyte clusters. J Cellular communication analysis revealing potential interactions among pericytes and other cell types. K Violin-box plots representing the expression of proinflammatory or anti-inflammatory signature genes in the macrophage clusters. Macrophage cells in control group, n = 6201; macrophage cells in MAT2A LOF group, n = 6763. L UMAP plot and quantitative analysis showing characterized cell clusters of infiltrated macrophages and their proportions in the indicated groups. M Cell trajectory analysis of the characterized cell clusters of infiltrated macrophages. N Representative immunofluorescence images and statistical analysis demonstrating the macrophageinfiltration in cutaneous wound tissues on day 8. n = 3 mice in each group. Scale bar, 20 μm. For the box and violin-box plots in ( G ), and ( K ), the centerlines indicated the medians. The box limits indicated the first and third quartiles. The whiskers indicated the maxima and minima. Data in the bar plots were shown as mean ± SD. n = 3 biologically independent samples ( D , G ). Statistical significance was determined using two-way ANOVA with Tukey’s multiple comparisons test ( C ) and two-tailed unpaired t test ( D , G , K , N ). Source data are provided as a Source Data file.

Article Snippet: D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration.

Techniques: Western Blot, Expressing, Isolation, Software, Sampling, Staining, Immunohistochemistry, Immunofluorescence, Control, Two Tailed Test

A Gene set enrichment analysis derived from the single-cell transcriptome profiling of the wound margin tissue showing the differential biological processes in the pericyte clusters between MAT2A LOF group and control group. NES, Nominal Enrichment Score. FDR P values were calculated based on the one-tailed test on the appropriate side of the null distribution. B Gene set variation analysis derived from the single-cell transcriptome profiling showing the significant enrichment of senescence-related pathway terms in the pericyte clusters. C Box plot showing the recommendation indices of six SIDs for the pericyte cluster ( n = 2211 cells), with SID3 presenting the highest values. D Box plot showing the SID3 scores of pericyte clusters between the MAT2A LOF ( n = 1025 cells) and control groups ( n = 1186 cells). E UMAP plots showing the distribution of SID3 scores of pericyte clusters in the indicated groups. F Immunostaining and statistical analysis of P21 (Red) and NG2 (Green) in cutaneous wound tissues between the control group and the MAT2A LOF group. Scale bar, 20 μm. G Effect of Mat2a knockdown on pericyte senescence, as determined by SAHF formation (H3K9me3 staining) and SA-β-gal staining. Pericytes were isolated from the mouse skins. H Western blot analysis determining the expression levels of P21 and Lamin B1 of pericytes in the indicated groups. I , J OCR measurement and maximal respiration analysis of pericytes with or without Mat2a knockdown. K Bar plot showing the cellular ATP levels of pericytes with or without Mat2a knockdown. L Bar plots showing MAT2A mRNA levels in cells from humans and mice. For the box plots in ( C ), and ( D ), the centerlines indicated the medians. The box limits indicated the first and third quartiles. The whiskers indicated the maxima and minima. Data in the bar plots were shown as mean ± SD. n = 3 biologically independent samples ( F , G , I , J , K ). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test ( G ) and two-tailed unpaired t test ( D , F , J , K ). Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: A biomimetic senotherapy replenishing MAT2A promotes wound regeneration in preclinical models

doi: 10.1038/s41467-025-65659-2

Figure Lengend Snippet: A Gene set enrichment analysis derived from the single-cell transcriptome profiling of the wound margin tissue showing the differential biological processes in the pericyte clusters between MAT2A LOF group and control group. NES, Nominal Enrichment Score. FDR P values were calculated based on the one-tailed test on the appropriate side of the null distribution. B Gene set variation analysis derived from the single-cell transcriptome profiling showing the significant enrichment of senescence-related pathway terms in the pericyte clusters. C Box plot showing the recommendation indices of six SIDs for the pericyte cluster ( n = 2211 cells), with SID3 presenting the highest values. D Box plot showing the SID3 scores of pericyte clusters between the MAT2A LOF ( n = 1025 cells) and control groups ( n = 1186 cells). E UMAP plots showing the distribution of SID3 scores of pericyte clusters in the indicated groups. F Immunostaining and statistical analysis of P21 (Red) and NG2 (Green) in cutaneous wound tissues between the control group and the MAT2A LOF group. Scale bar, 20 μm. G Effect of Mat2a knockdown on pericyte senescence, as determined by SAHF formation (H3K9me3 staining) and SA-β-gal staining. Pericytes were isolated from the mouse skins. H Western blot analysis determining the expression levels of P21 and Lamin B1 of pericytes in the indicated groups. I , J OCR measurement and maximal respiration analysis of pericytes with or without Mat2a knockdown. K Bar plot showing the cellular ATP levels of pericytes with or without Mat2a knockdown. L Bar plots showing MAT2A mRNA levels in cells from humans and mice. For the box plots in ( C ), and ( D ), the centerlines indicated the medians. The box limits indicated the first and third quartiles. The whiskers indicated the maxima and minima. Data in the bar plots were shown as mean ± SD. n = 3 biologically independent samples ( F , G , I , J , K ). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test ( G ) and two-tailed unpaired t test ( D , F , J , K ). Source data are provided as a Source Data file.

Article Snippet: D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration.

Techniques: Derivative Assay, Single Cell, Control, One-tailed Test, Immunostaining, Knockdown, Staining, Isolation, Western Blot, Expressing, Two Tailed Test

$A Bubble map showing the interactions of selected ligand-receptor pairs between pericytes and macrophage subsets. The communication strengths of interacting molecule 1 in cluster 1 and interacting molecule 2 in cluster 2 were indicated by color gradients (blue, low level; red, high level). The P values were calculated with the CellChat one-sided permutation test and indicated by the circle size. B Top panel: The coculture workflow of pericytes with or without Mat2a knockdown and macrophages (bone marrow-derived macrophages). Bottom panel: RT-qPCR analysis of inflammatory signature genes in macrophages. In the contact coculture system, the macrophages were collected through magnetic bead cell sorting (MACS). The workflow was created with BioRender.com. C Heatmap representing the change direction of senescence associated secretion phenotype in pericytes within the wound margin of the indicated groups. D In vitro RT-qPCR verifying the expression of a subset of typical SASP factors in pericytes with or without Mat2a knockdown. E Bar plots showing the Rcm values across a wide range of rank cutoffs (10%-100%) for pericytes within the wound margin of the MAT2A LOF group and the control group. The labels A1-A3 and B1-B3 represent the sample numbers within the groups. F Violin plots depicting the estimated strength (left) and fraction (right) of macrophage-derived mitochondria in pericytes predicted by MERCI between the MAT2A LOF group (n = 154 cells) and control group (n = 184 cells). G In vitro coculture of GFP+ pericytes and mitoDsRed+ macrophages immunostained with β-actin (white) to visualize the membrane boundaries of the two cell types, emphasized with high magnification. Arrows indicate transferred mitoDsRed+ mitochondria in pericytes. Scale bar, 10 μm.For the violin-box plots in ( F ), the centerlines indicated the medians. The box limits indicated the first and third quartiles. The whiskers indicated the maxima and minima. Data in the bar plots were shown as mean ± SD. n = 3 biologically independent samples ( B , D , G ). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test ( D ) and two-tailed unpaired t test ( B , F , G ). Source data are provided as a Source Data file.$

Journal: Nature Communications

Article Title: A biomimetic senotherapy replenishing MAT2A promotes wound regeneration in preclinical models

doi: 10.1038/s41467-025-65659-2

Figure Lengend Snippet: A Bubble map showing the interactions of selected ligand-receptor pairs between pericytes and macrophage subsets. The communication strengths of interacting molecule 1 in cluster 1 and interacting molecule 2 in cluster 2 were indicated by color gradients (blue, low level; red, high level). The P values were calculated with the CellChat one-sided permutation test and indicated by the circle size. B Top panel: The coculture workflow of pericytes with or without Mat2a knockdown and macrophages (bone marrow-derived macrophages). Bottom panel: RT-qPCR analysis of inflammatory signature genes in macrophages. In the contact coculture system, the macrophages were collected through magnetic bead cell sorting (MACS). The workflow was created with BioRender.com. C Heatmap representing the change direction of senescence associated secretion phenotype in pericytes within the wound margin of the indicated groups. D In vitro RT-qPCR verifying the expression of a subset of typical SASP factors in pericytes with or without Mat2a knockdown. E Bar plots showing the Rcm values across a wide range of rank cutoffs (10%-100%) for pericytes within the wound margin of the MAT2A LOF group and the control group. The labels A1-A3 and B1-B3 represent the sample numbers within the groups. F Violin plots depicting the estimated strength (left) and fraction (right) of macrophage-derived mitochondria in pericytes predicted by MERCI between the MAT2A LOF group (n = 154 cells) and control group (n = 184 cells). G In vitro coculture of GFP+ pericytes and mitoDsRed+ macrophages immunostained with β-actin (white) to visualize the membrane boundaries of the two cell types, emphasized with high magnification. Arrows indicate transferred mitoDsRed+ mitochondria in pericytes. Scale bar, 10 μm.For the violin-box plots in ( F ), the centerlines indicated the medians. The box limits indicated the first and third quartiles. The whiskers indicated the maxima and minima. Data in the bar plots were shown as mean ± SD. n = 3 biologically independent samples ( B , D , G ). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test ( D ) and two-tailed unpaired t test ( B , F , G ). Source data are provided as a Source Data file.

Article Snippet: D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration.

Techniques: Knockdown, Derivative Assay, Quantitative RT-PCR, FACS, In Vitro, Expressing, Control, Membrane, Two Tailed Test

A Effect of endogenous Mat2a knockdown followed by restoration of Mat2a WT, MUT1 or SAM (500 μM) on pericyte senescence, as determined by SAHF formation (H3K9me3 staining) and SA-β-gal staining ( n = 3). B Western blot assessment of the expression levels of P21 and Lamin B1 in the indicated pericytes. C – E OCR measurement, maximal respiration analysis and cellular ATP assessment of pericytes following endogenous Mat2a knockdown with Mat2a WT, MUT1 or SAM (500 μM) restoration ( n = 3). F Scheme displaying the procedure used for identifying the specific targets of MAT2A through proteomic and IP-MS analysis. The workflow was created with BioRender.com. G Heatmap showing the change direction of differential proteins in pericytes with or without Mat2a knockdown. H Display showed the differentially regulated proteins, categorized per known or predicted function(s), literature and sequence similarity. Circle size was proportional to the number of differentially expressed proteins. I Intersection of the results from the proteomics and IP-MS analyses. J Scheme displaying the HMGCS1-mediated MVA pathway. K IP and WB analyses showing the interaction of MAT2A and HMGCS1 in 293T cells with indicated transfections. L In vitro binding analysis of MAT2A and HMGCS1 with GST pull-down assays. M Design of MAT2A and HMGCS1 truncations. N IP and WB analysis representing the interactions between Flag-tagged truncated MAT2A and His-tagged PRMT1 proteins in 293T cells. O IP and WB analysis representing the interactions between His-tagged truncated HMGCS1 and Flag-tagged MAT2A proteins in 293T cells. P Molecular docking showing the interaction between MAT2A truncation (slate) and HMGCS1 truncation (cyan). Q Docked positions of MAT2A and HMGCS1 and design of the mutations of binding sites between MAT2A and HMGCS1. R IP and WB analysis of the interactions between FLAG-tagged MAT2A mutation (MUT2) and His-tagged HMGCS1 mutation in 293T cells. Data were shown as mean ± SD. n = 3 biologically independent samples ( A , C , D , E ). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test ( A , D , E ). n.s. no significance. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: A biomimetic senotherapy replenishing MAT2A promotes wound regeneration in preclinical models

doi: 10.1038/s41467-025-65659-2

Figure Lengend Snippet: A Effect of endogenous Mat2a knockdown followed by restoration of Mat2a WT, MUT1 or SAM (500 μM) on pericyte senescence, as determined by SAHF formation (H3K9me3 staining) and SA-β-gal staining ( n = 3). B Western blot assessment of the expression levels of P21 and Lamin B1 in the indicated pericytes. C – E OCR measurement, maximal respiration analysis and cellular ATP assessment of pericytes following endogenous Mat2a knockdown with Mat2a WT, MUT1 or SAM (500 μM) restoration ( n = 3). F Scheme displaying the procedure used for identifying the specific targets of MAT2A through proteomic and IP-MS analysis. The workflow was created with BioRender.com. G Heatmap showing the change direction of differential proteins in pericytes with or without Mat2a knockdown. H Display showed the differentially regulated proteins, categorized per known or predicted function(s), literature and sequence similarity. Circle size was proportional to the number of differentially expressed proteins. I Intersection of the results from the proteomics and IP-MS analyses. J Scheme displaying the HMGCS1-mediated MVA pathway. K IP and WB analyses showing the interaction of MAT2A and HMGCS1 in 293T cells with indicated transfections. L In vitro binding analysis of MAT2A and HMGCS1 with GST pull-down assays. M Design of MAT2A and HMGCS1 truncations. N IP and WB analysis representing the interactions between Flag-tagged truncated MAT2A and His-tagged PRMT1 proteins in 293T cells. O IP and WB analysis representing the interactions between His-tagged truncated HMGCS1 and Flag-tagged MAT2A proteins in 293T cells. P Molecular docking showing the interaction between MAT2A truncation (slate) and HMGCS1 truncation (cyan). Q Docked positions of MAT2A and HMGCS1 and design of the mutations of binding sites between MAT2A and HMGCS1. R IP and WB analysis of the interactions between FLAG-tagged MAT2A mutation (MUT2) and His-tagged HMGCS1 mutation in 293T cells. Data were shown as mean ± SD. n = 3 biologically independent samples ( A , C , D , E ). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test ( A , D , E ). n.s. no significance. Source data are provided as a Source Data file.

Article Snippet: D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration.

Techniques: Knockdown, Staining, Western Blot, Expressing, Protein-Protein interactions, Sequencing, Transfection, In Vitro, Binding Assay, Mutagenesis

A HMGCS1 expression levels in pericytes with Mat2a knockdown followed by transfection of Mat2a WT or MUT2. B HMGCS1 expression levels in pericytes treated with cycloheximide (CHX, 100 μg/ml) for the indicated times (top) and relative HMGCS1 protein levels (bottom). C HMGCS1 expression levels in Mat2a -knockdown pericytes treated with or without 10 μM MG132 for 8 h. D Ubiquitination of HMGCS1 in pericytes with the indicated transfections and treatment with 10 μM MG132 for 8 h. E Identification of ubiquitination related modification factors from IP/MS data. HMGCS1 expression levels in pericytes following Otub1 knockdown with or without WT restoration. The workflow was created with BioRender.com. F Ubiquitination of HMGCS1 in 293T cells with transfection of OTUB1 or the indicated mutant and treatment with 10 μM MG132 for 8 h. G Co-localization analysis of MAT2A, OTUB1 and HMGCS1 by immunofluorescence staining in pericytes. H Binding analysis of HMGCS1 and OTUB1 following MAT2A transfection or not in 293T cells treated with 10 μM MG132 for 8 h. I Ubiquitination of HMGCS1 in 293T cells with the indicated transfection of OTUB1 and MAT2A and treatment with 10 μM MG132 for 8 h. J Binding analysis of HMGCS1 and OTUB1 following MAT2A knockdown or not in 293T cells treated with 10 μM MG132 for 8 h. K Ubiquitination of HMGCS1 in 293T cells with the indicated transfection of OTUB1 and knockdown of MAT2A and treatment with 10 μM MG132 for 8 h. L Co-localization analysis of OTUB1 and HMGCS1 following Mat2a knockdown by immunofluorescence staining in pericytes. Data were shown as mean ± SD. n = 3 biologically independent samples ( B ). Statistical significance was determined using two-way ANOVA with Tukey’s multiple comparisons test ( B ). n.s. no significance. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: A biomimetic senotherapy replenishing MAT2A promotes wound regeneration in preclinical models

doi: 10.1038/s41467-025-65659-2

Figure Lengend Snippet: A HMGCS1 expression levels in pericytes with Mat2a knockdown followed by transfection of Mat2a WT or MUT2. B HMGCS1 expression levels in pericytes treated with cycloheximide (CHX, 100 μg/ml) for the indicated times (top) and relative HMGCS1 protein levels (bottom). C HMGCS1 expression levels in Mat2a -knockdown pericytes treated with or without 10 μM MG132 for 8 h. D Ubiquitination of HMGCS1 in pericytes with the indicated transfections and treatment with 10 μM MG132 for 8 h. E Identification of ubiquitination related modification factors from IP/MS data. HMGCS1 expression levels in pericytes following Otub1 knockdown with or without WT restoration. The workflow was created with BioRender.com. F Ubiquitination of HMGCS1 in 293T cells with transfection of OTUB1 or the indicated mutant and treatment with 10 μM MG132 for 8 h. G Co-localization analysis of MAT2A, OTUB1 and HMGCS1 by immunofluorescence staining in pericytes. H Binding analysis of HMGCS1 and OTUB1 following MAT2A transfection or not in 293T cells treated with 10 μM MG132 for 8 h. I Ubiquitination of HMGCS1 in 293T cells with the indicated transfection of OTUB1 and MAT2A and treatment with 10 μM MG132 for 8 h. J Binding analysis of HMGCS1 and OTUB1 following MAT2A knockdown or not in 293T cells treated with 10 μM MG132 for 8 h. K Ubiquitination of HMGCS1 in 293T cells with the indicated transfection of OTUB1 and knockdown of MAT2A and treatment with 10 μM MG132 for 8 h. L Co-localization analysis of OTUB1 and HMGCS1 following Mat2a knockdown by immunofluorescence staining in pericytes. Data were shown as mean ± SD. n = 3 biologically independent samples ( B ). Statistical significance was determined using two-way ANOVA with Tukey’s multiple comparisons test ( B ). n.s. no significance. Source data are provided as a Source Data file.

Article Snippet: D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration.

Techniques: Expressing, Knockdown, Transfection, Ubiquitin Proteomics, Modification, Protein-Protein interactions, Mutagenesis, Immunofluorescence, Staining, Binding Assay

A Cellular CoQ levels in pericytes with Mat2a knockdown. B MAT2A and HMGCS1 expression levels in pericytes with the indicated transfections. C Cellular CoQ levels in pericytes with Mat2a knockdown followed by transfection of Mat2a MUT2, Hmgcs1 , or not. D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration. E , F ATP assessment and cell viability in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ restoration. G SA-β-gal staining in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ restoration. H Expression levels of P21 and Lamin B1 in pericytes subjected to the indicated treatments. I RT-qPCR analysis showing the expression levels of typical SASP components in pericytes subjected to the indicated treatments. J , K MitoDsRed+ macrophages were co-cultured with GFP+ pericytes following the indicated treatment. GFP+ MitoDsRed+ pericytes were quantified via flow cytometry ( J ), as summarized in ( K ).Data were shown as mean ± SD. n = 3 biologically independent samples ( A , C – G , I , K ). Statistical significance was determined using two-tailed unpaired t test ( A ), one-way ANOVA with Tukey’s multiple comparisons test ( C – E , G , I , K ) and two-way ANOVA with Tukey’s multiple comparisons test ( F ). n.s. no significance. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: A biomimetic senotherapy replenishing MAT2A promotes wound regeneration in preclinical models

doi: 10.1038/s41467-025-65659-2

Figure Lengend Snippet: A Cellular CoQ levels in pericytes with Mat2a knockdown. B MAT2A and HMGCS1 expression levels in pericytes with the indicated transfections. C Cellular CoQ levels in pericytes with Mat2a knockdown followed by transfection of Mat2a MUT2, Hmgcs1 , or not. D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration. E , F ATP assessment and cell viability in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ restoration. G SA-β-gal staining in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ restoration. H Expression levels of P21 and Lamin B1 in pericytes subjected to the indicated treatments. I RT-qPCR analysis showing the expression levels of typical SASP components in pericytes subjected to the indicated treatments. J , K MitoDsRed+ macrophages were co-cultured with GFP+ pericytes following the indicated treatment. GFP+ MitoDsRed+ pericytes were quantified via flow cytometry ( J ), as summarized in ( K ).Data were shown as mean ± SD. n = 3 biologically independent samples ( A , C – G , I , K ). Statistical significance was determined using two-tailed unpaired t test ( A ), one-way ANOVA with Tukey’s multiple comparisons test ( C – E , G , I , K ) and two-way ANOVA with Tukey’s multiple comparisons test ( F ). n.s. no significance. Source data are provided as a Source Data file.

Article Snippet: D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration.

Techniques: Knockdown, Expressing, Transfection, Staining, Quantitative RT-PCR, Cell Culture, Flow Cytometry, Two Tailed Test

Harmonious cellular communication and cooperation, as well as efficient transformation of cell phenotypes, are indispensable for wound regeneration. MAT2A downregulation mediated pericyte senescence in a moonlighting manner, which induced the infiltration of inflammatory macrophages in diabetic wounds. This discovery provides an saRNA-based strategy targeting senescent pericytes for wound healing. The diagram was created with BioRender.com.

Journal: Nature Communications

Article Title: A biomimetic senotherapy replenishing MAT2A promotes wound regeneration in preclinical models

doi: 10.1038/s41467-025-65659-2

Figure Lengend Snippet: Harmonious cellular communication and cooperation, as well as efficient transformation of cell phenotypes, are indispensable for wound regeneration. MAT2A downregulation mediated pericyte senescence in a moonlighting manner, which induced the infiltration of inflammatory macrophages in diabetic wounds. This discovery provides an saRNA-based strategy targeting senescent pericytes for wound healing. The diagram was created with BioRender.com.

Article Snippet: D OCR measurement in pericytes with Mat2a knockdown followed by transfection of Hmgcs1 , or CoQ (TargetMol, USA, T2796) restoration.

Techniques: Transformation Assay

a Western blots show the time course of protein expression of FOXM1, MATα2, and MAT2β in liver tissues after BDL ( n = 3 independent experiments). b Immunofluorescence (IF) of FOXM1, MATα2, and MAT2β in primary hepatic stellate cells (HSCs) isolated from sham and BDL mice at day 5. The top row shows DAPI staining. The second and third rows show the antibody (AB) staining. The fourth row shows merged images of DAPI and FOXM1, MATα2 or MAT2β, and the fifth row shows high magnification (HM) from the merged image ( n = 3 independent experiments). c Expression of mRNA (top) and protein (bottom) of FOXM1, MATα2, MAT2β, α-SMA, and COL1A1 in HSCs isolated from WT mice and cultured for up to 5 days and FDI-6 treatment for 24 h starting at day 4. Data presented as mean ± SEM ( n = 3 per group), mRNA levels of Foxm1, Mat2a, Mat2b , Acta2 and Col1a1 in HSCs at day 5 vs. day 1, p = 0.0044, p = 0.0029, p = 0.0056, p = 0.0010 and p = 0.00002, respectively. mRNA levels of Foxm1, Mat2a, Mat2b , Acta2 and Col1a1 in HSCs at day 5 + FDI-6 vs. day 1, p = 0.0431, p = 0.0152, p = 0.0245, p = 0.0083, and p = 0.0082, respectively. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. See Supplementary Fig. for densitometric values of the western blots. d Expression of mRNA and protein of FOXM1, MATα2, MAT2β, α-SMA, and COL1A1 after FDI-6 treatment in LX-2 cells. Data presented as mean ± SEM ( n = 3 per group), mRNA levels of Foxm1, Mat2a, Mat2b , Acta2 and Col1a1 in LX2 cells with DMSO treatment vs. FDI-6, p = 0.0187, p = 0.0023, p = 0.0122, p = 0.0108 and p = 0.0124, respectively. * p < 0.05, ** p < 0.01 vs. DMSO. See Supplementary Fig. for densitometric values of the western blots. e IF of LX-2 cells after treatment with FDI-6. HM, high magnification from the merged images ( n = 3 independent experiments). f FOXM1, MATα2, and MAT2β in cytoplasm and nucleus from HSCs isolated from sham and BDL mice with or without FDI-6 treatment ( n = 3 independent experiments). Densitometry for cytoplasmic protein levels is summarized in Supplementary Fig. and nuclear protein levels is summarized in Supplementary Fig. . Proliferation ( g ) and migration ( h ) of LX-2 cells in vitro after FDI-6 treatment for 24 h. Data presented as mean ± SEM ( n = 3 per group). p = 0.00016, p = 0.00002 vs.DMSO. Statistical significance was determined by using two-tailed unpaired Student’s t -test. *** p < 0.001, **** p < 0.0001 vs. DMSO ( n = 3). Abbreviations: BDL bile duct ligation, DMSO dimethylsulfoxide. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: The role of forkhead box M1-methionine adenosyltransferase 2 A/2B axis in liver inflammation and fibrosis

doi: 10.1038/s41467-024-52527-8

Figure Lengend Snippet: a Western blots show the time course of protein expression of FOXM1, MATα2, and MAT2β in liver tissues after BDL ( n = 3 independent experiments). b Immunofluorescence (IF) of FOXM1, MATα2, and MAT2β in primary hepatic stellate cells (HSCs) isolated from sham and BDL mice at day 5. The top row shows DAPI staining. The second and third rows show the antibody (AB) staining. The fourth row shows merged images of DAPI and FOXM1, MATα2 or MAT2β, and the fifth row shows high magnification (HM) from the merged image ( n = 3 independent experiments). c Expression of mRNA (top) and protein (bottom) of FOXM1, MATα2, MAT2β, α-SMA, and COL1A1 in HSCs isolated from WT mice and cultured for up to 5 days and FDI-6 treatment for 24 h starting at day 4. Data presented as mean ± SEM ( n = 3 per group), mRNA levels of Foxm1, Mat2a, Mat2b , Acta2 and Col1a1 in HSCs at day 5 vs. day 1, p = 0.0044, p = 0.0029, p = 0.0056, p = 0.0010 and p = 0.00002, respectively. mRNA levels of Foxm1, Mat2a, Mat2b , Acta2 and Col1a1 in HSCs at day 5 + FDI-6 vs. day 1, p = 0.0431, p = 0.0152, p = 0.0245, p = 0.0083, and p = 0.0082, respectively. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. See Supplementary Fig. for densitometric values of the western blots. d Expression of mRNA and protein of FOXM1, MATα2, MAT2β, α-SMA, and COL1A1 after FDI-6 treatment in LX-2 cells. Data presented as mean ± SEM ( n = 3 per group), mRNA levels of Foxm1, Mat2a, Mat2b , Acta2 and Col1a1 in LX2 cells with DMSO treatment vs. FDI-6, p = 0.0187, p = 0.0023, p = 0.0122, p = 0.0108 and p = 0.0124, respectively. * p < 0.05, ** p < 0.01 vs. DMSO. See Supplementary Fig. for densitometric values of the western blots. e IF of LX-2 cells after treatment with FDI-6. HM, high magnification from the merged images ( n = 3 independent experiments). f FOXM1, MATα2, and MAT2β in cytoplasm and nucleus from HSCs isolated from sham and BDL mice with or without FDI-6 treatment ( n = 3 independent experiments). Densitometry for cytoplasmic protein levels is summarized in Supplementary Fig. and nuclear protein levels is summarized in Supplementary Fig. . Proliferation ( g ) and migration ( h ) of LX-2 cells in vitro after FDI-6 treatment for 24 h. Data presented as mean ± SEM ( n = 3 per group). p = 0.00016, p = 0.00002 vs.DMSO. Statistical significance was determined by using two-tailed unpaired Student’s t -test. *** p < 0.001, **** p < 0.0001 vs. DMSO ( n = 3). Abbreviations: BDL bile duct ligation, DMSO dimethylsulfoxide. Source data are provided as a Source Data file.

Article Snippet: Human and mouse probes for MAT2A , MAT2B and FOXM1 , and the PCR Supermix were purchased from ThermoFisher.

Techniques: Western Blot, Expressing, Immunofluorescence, Isolation, Staining, Cell Culture, Migration, In Vitro, Two Tailed Test, Ligation

a Liver sections from prevention groups of the corn oil (Oil) + DMSO, Oil + FDI-6, CCl 4 + DMSO, and CCl 4 + FDI-6 for three weeks, and treatment groups of Oil + DMSO, Oil + FDI-6, CCl 4 + DMSO, and CCl 4 + FDI-6 treated for two weeks after CCl 4 treatment for three weeks. IHC stained with antibodies of COL1A1, F4/80, α–SMA, FOXM1, MATα2, and MAT2β. H&E is shown in the top row. b , c show changes in ALT ( n = 6 per group) ( b ) and AST ( n = 6 in prevention group of Oil + DMSO, n = 5 in prevention group of Oil + FDI-6, n = 3 in prevention group of CCl 4 + DMSO and treatment group of Oil + FDI-6, n = 4 in prevention group of CCl 4 + FDI-6, and treatment groups of Oil + DMSO, CCl 4 + DMSO, and CCl 4 + FDI-6) ( c ) levels after FDI-6 administration in the prevention and treatment groups. Data presented as mean ± SEM, **p < 0.01, **** p < 0.0001. d Hydroxyproline content was measured in the livers from prevention and treatment groups with or without FDI-6 administration. Data presented as mean ± SEM, **p < 0.01, **** p < 0.0001 ( n = 5 per group). e mRNA levels of Foxm1 , Mat2a , and Mat2b in the livers after FDI-6 administration in the prevention and treatment groups. Data presented as mean ± SEM ( n = 4), **p < 0.01, **** p < 0.0001 ( n = 4 in prevention and treatment groups of Oil + DMSO for Foxm1 mRNA; n = 6 in prevention and treatment groups of Oil + DMSO for Mat2α and Mat2b mRNA; n = 4 in prevention and treatment groups of Oil+ FDI-6 for Foxm1 mRNA; n = 6 in prevention and treatment groups of Oil + FDI-6 for Mat2α and Mat2b mRNA level; n = 4 in prevention group of CCl 4 + DMSO for Foxm1 mRNA level; n = 6 in prevention and treatment groups of CCl 4 + DMSO for Mat2α and Mat2b mRNA and treatment group of CCl 4 + DMSO for Foxm1 mRNA; n = 5 in treatment group of CCl 4 + FDI-6 for Foxm1 mRNA level; n = 6 in prevention and treatment groups of CCl 4 + FDI-6 for Mat2α and Mat2b mRNA and prevention group of CCl4+FDI-6 for Foxm1 mRNA. f Protein levels of FOXM1, MATα2, MAT2β, α–SMA, F4/80, and COL1A1 from livers after prevention and treatment with FDI-6 ( n = 3 independent experiments). Densitometry values for protein levels are summarized in Supplementary Fig. . Statistical significance was determined by using two-tailed, unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: ALT alanine transaminase, AST aspartate aminotransferase.

Journal: Nature Communications

Article Title: The role of forkhead box M1-methionine adenosyltransferase 2 A/2B axis in liver inflammation and fibrosis

doi: 10.1038/s41467-024-52527-8

Figure Lengend Snippet: a Liver sections from prevention groups of the corn oil (Oil) + DMSO, Oil + FDI-6, CCl 4 + DMSO, and CCl 4 + FDI-6 for three weeks, and treatment groups of Oil + DMSO, Oil + FDI-6, CCl 4 + DMSO, and CCl 4 + FDI-6 treated for two weeks after CCl 4 treatment for three weeks. IHC stained with antibodies of COL1A1, F4/80, α–SMA, FOXM1, MATα2, and MAT2β. H&E is shown in the top row. b , c show changes in ALT ( n = 6 per group) ( b ) and AST ( n = 6 in prevention group of Oil + DMSO, n = 5 in prevention group of Oil + FDI-6, n = 3 in prevention group of CCl 4 + DMSO and treatment group of Oil + FDI-6, n = 4 in prevention group of CCl 4 + FDI-6, and treatment groups of Oil + DMSO, CCl 4 + DMSO, and CCl 4 + FDI-6) ( c ) levels after FDI-6 administration in the prevention and treatment groups. Data presented as mean ± SEM, **p < 0.01, **** p < 0.0001. d Hydroxyproline content was measured in the livers from prevention and treatment groups with or without FDI-6 administration. Data presented as mean ± SEM, **p < 0.01, **** p < 0.0001 ( n = 5 per group). e mRNA levels of Foxm1 , Mat2a , and Mat2b in the livers after FDI-6 administration in the prevention and treatment groups. Data presented as mean ± SEM ( n = 4), **p < 0.01, **** p < 0.0001 ( n = 4 in prevention and treatment groups of Oil + DMSO for Foxm1 mRNA; n = 6 in prevention and treatment groups of Oil + DMSO for Mat2α and Mat2b mRNA; n = 4 in prevention and treatment groups of Oil+ FDI-6 for Foxm1 mRNA; n = 6 in prevention and treatment groups of Oil + FDI-6 for Mat2α and Mat2b mRNA level; n = 4 in prevention group of CCl 4 + DMSO for Foxm1 mRNA level; n = 6 in prevention and treatment groups of CCl 4 + DMSO for Mat2α and Mat2b mRNA and treatment group of CCl 4 + DMSO for Foxm1 mRNA; n = 5 in treatment group of CCl 4 + FDI-6 for Foxm1 mRNA level; n = 6 in prevention and treatment groups of CCl 4 + FDI-6 for Mat2α and Mat2b mRNA and prevention group of CCl4+FDI-6 for Foxm1 mRNA. f Protein levels of FOXM1, MATα2, MAT2β, α–SMA, F4/80, and COL1A1 from livers after prevention and treatment with FDI-6 ( n = 3 independent experiments). Densitometry values for protein levels are summarized in Supplementary Fig. . Statistical significance was determined by using two-tailed, unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: ALT alanine transaminase, AST aspartate aminotransferase.

Article Snippet: Human and mouse probes for MAT2A , MAT2B and FOXM1 , and the PCR Supermix were purchased from ThermoFisher.

Techniques: Staining, Two Tailed Test

MAT2A ( a ), MAT2B ( b ) and FOXM1 ( c ) promoter activities in LX-2 cells and primary cholangiocytes ± FDI-6 or siRNA treatment as described in Methods. Effects of mutating FOX elements in LX-2 cells are shown in ( d ) MAT2A , ( e ) MAT2B , and ( f ) FOXM1 . Cells transfected with WT and mutant constructs were treated with siRNA against MAT2A, MAT2B, and FOXM1 and reporter activities were measured. Data presented as mean ± SEM ( n = 3 per group). For a, Mat2a promoter activities of D-271/ + 60, D-671/ + 60 and D-1329/ + 60 in LX2 cell, SC + DMSO vs. SC + FDI-6, p = 0.0002, p = 0.0052 and p = 0.0948, respectively; SC + DMASO vs. SC + FOXM1 si, p = 0.0001, p = 0.0022 and p = 0.0003, respectively; Mat2a promoter activities of D-271/ + 60 in cholangiocytes, SC + DMSO vs. SC + FDI-6 or SC+Foxm1si, p = 0.0286 and 0.0149, respectively. For b , Mat2b promoter activities of D-25−/+3, D-713/+3, D-990/+3 and D-1319/+3 in LX2 cell, SC + DMSO vs. SC + FDI-6, p = 0.0051, p = 0.0316, p = 0.016 and p = 0.0034, respectively; SC + DMSO vs. SC + FOXM1 si, p = 0.0106, p = 0.0233, p = 0.0019 and p = 0.0049 respectively; Mat2b promoter activities of D-250/+3 in cholangiocytes, SC + DMSO vs. SC + FDI-6 or SC + Foxm1 si, p = 0.0002 and 0.023, respectively. For c , Foxm1 promoter activities of D-312/+107 and D-1333/+107 in LX2 cell, SC + DMSO vs. SC + FDI-6, p = 0.5799 and p = 0.0020, respectively; SC + DMSO vs. SC + FOXM1 si, p = 0.7684 and p = 0.0047 respectively; Foxm1 promoter activities of D-1333/ + 107 in cholangiocytes, SC + DMSO vs. SC + FDI-6 or SC + Foxm1 si, p = 0.0163 and 0.0131, respectively. For d , MAT2A promoter activities (−270/+60) of WT and MU, SC vs. SC, p = 0.015 and p = 0.28 respectively; FOXM1si vs. SC, p = 0.0032 and p = 0.042 respectively; MAT2Asi vs. SC, p = 0.0033 and p = 0.066 respectively. For e , MAT2B promoter activities (−250/+3) of WT and MU, SC vs. SC, p = 0.0017 and p = 0.11 respectively; FOXM1si vs. SC, p = 0.0018 and p = 0.022 respectively; MAT2Asi vs. SC, p = 0.0015 and p = 0.026 respectively. For f , FOXM1 promoter activities (−1333/+107) of WT and MU, SC vs. SC, p = 0.000017 and p = 0.0078 respectively; FOXM1si vs. SC, p = 0.000014 and p = 0.0063 respectively; MAT2Asi vs. SC, p = 0.00020 and p = 0.019 respectively. g ChIP assay was performed by spanning two FOX regions of the FOXM1 promoter in LX-2 cells using FOXM1, MATα2 and MAT2β antibodies after treatments that varied the expression of FOXM1, MAT2A or MAT2B in the top three rows. Seq-ChIP with anti-MATα2 and MAT2β antibodies after FOXM1 ChIP was performed as described in Methods. Representative results from three experiments are shown. h qPCR analysis of the ChIP assay from ( g ). For h , ChIP and Seq-ChIP percentage of input DNA, FOXM1 si vs. SC, MAT2A si vs. SC, MAT2B si vs. SC, FOXM1 OV vs. EV, MAT2A OV vs. EV, and MAT2B OV vs. EV, for FOXM1 ChIP, p = 0.00089, p = 0.011, p = 0.0045, p = 0.0000053, p = 0.00024, and p = 0.000039; for MAT2A seq-ChIP p = 0.0015, p = 0.0023, p = 0.0037, p = 0.000037, p = 0.00076, and p = 0.00032; for MAT2B seq-ChIP p = 0.0036, p = 0.018, p = 0.0092, p = 0.0036, p = 0.0000098, p = 0.0013 respectively. Data presented as mean ± SEM, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. SC or EV ( n = 3 independent experiments). i EMSA was done using labeled probes containing two FOX binding motifs of the FOXM1 promoter as shown in ( f ) and 100 ug of nuclear protein from LX-2 cells after treatments that varied FOXM1/MAT2A/MAT2B ( n = 3 independent experiments). j Super shifts were done using 100 ng of recombinant proteins of FOXM1, MATα2, MAT2β alone or combined, and antibodies to FOXM1, MATα2 and MAT2β. Probe and IgG only served as negative controls. Results represent three independent experiments. k In vitro pull-down shows direct interaction between MATα2, MAT2β and FOXM1 using recombinant MATα2, MAT2β and FOXM1 proteins ( n = 3 independent experiments). l MATα2, MAT2β and FOXM1 interaction in Flox control (WT), Foxm1 Hep−/− , with or without BDL was detected by Co-IP and western blotting ( n = 3 independent experiments). Statistical significance was determined by using two-tailed, unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: AB antibody, EV empty vector, IP immunoprecipitation, OV overexpression, si siRNA, WT wild type, MU mutants.

Journal: Nature Communications

Article Title: The role of forkhead box M1-methionine adenosyltransferase 2 A/2B axis in liver inflammation and fibrosis

doi: 10.1038/s41467-024-52527-8

Figure Lengend Snippet: MAT2A ( a ), MAT2B ( b ) and FOXM1 ( c ) promoter activities in LX-2 cells and primary cholangiocytes ± FDI-6 or siRNA treatment as described in Methods. Effects of mutating FOX elements in LX-2 cells are shown in ( d ) MAT2A , ( e ) MAT2B , and ( f ) FOXM1 . Cells transfected with WT and mutant constructs were treated with siRNA against MAT2A, MAT2B, and FOXM1 and reporter activities were measured. Data presented as mean ± SEM ( n = 3 per group). For a, Mat2a promoter activities of D-271/ + 60, D-671/ + 60 and D-1329/ + 60 in LX2 cell, SC + DMSO vs. SC + FDI-6, p = 0.0002, p = 0.0052 and p = 0.0948, respectively; SC + DMASO vs. SC + FOXM1 si, p = 0.0001, p = 0.0022 and p = 0.0003, respectively; Mat2a promoter activities of D-271/ + 60 in cholangiocytes, SC + DMSO vs. SC + FDI-6 or SC+Foxm1si, p = 0.0286 and 0.0149, respectively. For b , Mat2b promoter activities of D-25−/+3, D-713/+3, D-990/+3 and D-1319/+3 in LX2 cell, SC + DMSO vs. SC + FDI-6, p = 0.0051, p = 0.0316, p = 0.016 and p = 0.0034, respectively; SC + DMSO vs. SC + FOXM1 si, p = 0.0106, p = 0.0233, p = 0.0019 and p = 0.0049 respectively; Mat2b promoter activities of D-250/+3 in cholangiocytes, SC + DMSO vs. SC + FDI-6 or SC + Foxm1 si, p = 0.0002 and 0.023, respectively. For c , Foxm1 promoter activities of D-312/+107 and D-1333/+107 in LX2 cell, SC + DMSO vs. SC + FDI-6, p = 0.5799 and p = 0.0020, respectively; SC + DMSO vs. SC + FOXM1 si, p = 0.7684 and p = 0.0047 respectively; Foxm1 promoter activities of D-1333/ + 107 in cholangiocytes, SC + DMSO vs. SC + FDI-6 or SC + Foxm1 si, p = 0.0163 and 0.0131, respectively. For d , MAT2A promoter activities (−270/+60) of WT and MU, SC vs. SC, p = 0.015 and p = 0.28 respectively; FOXM1si vs. SC, p = 0.0032 and p = 0.042 respectively; MAT2Asi vs. SC, p = 0.0033 and p = 0.066 respectively. For e , MAT2B promoter activities (−250/+3) of WT and MU, SC vs. SC, p = 0.0017 and p = 0.11 respectively; FOXM1si vs. SC, p = 0.0018 and p = 0.022 respectively; MAT2Asi vs. SC, p = 0.0015 and p = 0.026 respectively. For f , FOXM1 promoter activities (−1333/+107) of WT and MU, SC vs. SC, p = 0.000017 and p = 0.0078 respectively; FOXM1si vs. SC, p = 0.000014 and p = 0.0063 respectively; MAT2Asi vs. SC, p = 0.00020 and p = 0.019 respectively. g ChIP assay was performed by spanning two FOX regions of the FOXM1 promoter in LX-2 cells using FOXM1, MATα2 and MAT2β antibodies after treatments that varied the expression of FOXM1, MAT2A or MAT2B in the top three rows. Seq-ChIP with anti-MATα2 and MAT2β antibodies after FOXM1 ChIP was performed as described in Methods. Representative results from three experiments are shown. h qPCR analysis of the ChIP assay from ( g ). For h , ChIP and Seq-ChIP percentage of input DNA, FOXM1 si vs. SC, MAT2A si vs. SC, MAT2B si vs. SC, FOXM1 OV vs. EV, MAT2A OV vs. EV, and MAT2B OV vs. EV, for FOXM1 ChIP, p = 0.00089, p = 0.011, p = 0.0045, p = 0.0000053, p = 0.00024, and p = 0.000039; for MAT2A seq-ChIP p = 0.0015, p = 0.0023, p = 0.0037, p = 0.000037, p = 0.00076, and p = 0.00032; for MAT2B seq-ChIP p = 0.0036, p = 0.018, p = 0.0092, p = 0.0036, p = 0.0000098, p = 0.0013 respectively. Data presented as mean ± SEM, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. SC or EV ( n = 3 independent experiments). i EMSA was done using labeled probes containing two FOX binding motifs of the FOXM1 promoter as shown in ( f ) and 100 ug of nuclear protein from LX-2 cells after treatments that varied FOXM1/MAT2A/MAT2B ( n = 3 independent experiments). j Super shifts were done using 100 ng of recombinant proteins of FOXM1, MATα2, MAT2β alone or combined, and antibodies to FOXM1, MATα2 and MAT2β. Probe and IgG only served as negative controls. Results represent three independent experiments. k In vitro pull-down shows direct interaction between MATα2, MAT2β and FOXM1 using recombinant MATα2, MAT2β and FOXM1 proteins ( n = 3 independent experiments). l MATα2, MAT2β and FOXM1 interaction in Flox control (WT), Foxm1 Hep−/− , with or without BDL was detected by Co-IP and western blotting ( n = 3 independent experiments). Statistical significance was determined by using two-tailed, unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: AB antibody, EV empty vector, IP immunoprecipitation, OV overexpression, si siRNA, WT wild type, MU mutants.

Article Snippet: Human and mouse probes for MAT2A , MAT2B and FOXM1 , and the PCR Supermix were purchased from ThermoFisher.

Techniques: Transfection, Mutagenesis, Construct, Expressing, Labeling, Binding Assay, Recombinant, In Vitro, Control, Co-Immunoprecipitation Assay, Western Blot, Two Tailed Test, Plasmid Preparation, Immunoprecipitation, Over Expression

a H&E, Sirius red, CK19, α-SMA, and F4/80 staining in Flox control and Foxm1 Hep−/− mice after BDL as compared to sham surgery. Liver fibrosis was measured by Sirius red staining ( n = 7 per group) ( b ) and hydroxyproline assay ( n = 5 per group) ( c ), liver injury by ALT ( n = 6 per group) ( d ) and AST ( n = 6 per group) ( e ) levels, macrophage number by F4/80 ( n = 6 per group) ( f ), and ductular proliferation by CK19 ( n = 4 per group) and myofibroblast differentiation by α-SMA staining ( n = 3 per group) ( g ). For b , Sirus red area/total area for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.00000000011 and p = 0.0000015, respectively. For c , hydroxyproline (ug/g, liver) for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.000058 and p = 0.0034, respectively. For d , ALT level (ug/L) for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.000000000014 and p = 0.00000000097, respectively. For e , AST level (ug/L) for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.0000000000001 and p = 0.00000025, respectively. For f , F4/80 positive number for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.0000000020 and p = 0.00080, respectively. For g , CK19/total area for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.00000078 and p = 0.00026, respectively. Data are shown as mean ± SEM, ** p < 0.01, *** p < 0.001, **** p < 0.0001. h Protein expression of FOXM1, MATα2, and MAT2β in hepatocytes, cholangiocytes, HSCs, and KCs isolated from Flox control and Foxm1 Hep−/− mice ± BDL, Densitometry values for protein levels are summarized in Supplementary Fig. . mRNA levels of Foxm1 , Mat2a , and Mat2b in hepatocytes ( n = 6 animals in Flox + Sham, Foxm1 hep−/− + Sham, Flox + BDL groups and Foxm1 hep−/− + BDL group for foxm1 and mat2b mRNA; n = 4 in Flox + Sham, Foxm1 hep−/− + Sham, Flox + BDL groups and n = 6 in foxm1 hep−/− +BDL group for mat2a mRNA) ( i ), cholangiocytes ( n = 4 animals in Flox + Sham , foxm1 hep−/− + Sham, Flox + BDL groups and foxm1hep−/− + BDL groups for foxm1 mRNA; n = 5 in Flox + Sham and foxm1 hep−/− + Sham groups and n = 4 in Flox + BDL and Foxm1 hep-−/− + BDL groups for mat2a mRNA; n = 6 in Flox + Sham, Foxm1 hep−/− + Sham, Flox + BDL groups and foxm1 hep−/− + BDL group for mat2b mRNA) ( j ), HSCs ( n = 6 animals) ( k ), and KCs ( n = 4 animals in Flox + Sham, Flox + BDL groups, foxm1 hep−/− + BDL groups and n = 5 in foxm1 hep−/− + Sham for foxm1 mRNA; n = 5 in Flox + Sham and foxm1 hep−/− + Sham groups, Flox + BDL and Foxm1 hep−/− + BDL groups for mat2a mRNA; n = 5 in Flox + Sham, Flox + BDL and Foxm1 hep−/− + BDL groups and n = 6 in foxm1 hep−/− + Sham group for mat2b mRNA). l isolated from Flox control, Foxm1 Hep−/− mice ± BDL. Data are shown as mean fold of Flox control ± SEM, * p < 0.05, *** p < 0.001, **** p < 0.0001. p values obtained via two-tailed unpaired Student’s t tests. For i , mRNA levels in hepatocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham p = 0.0000045, p = 0.000030, and p = 0.000028 respectively; of Hep −/− BDL vs. Flox BDL p = 0.0000000099, p = 0.00024, p = 0.000099 respectively. For j , mRNA levels in cholangiocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham p = 0.00023, p = 0.00000026, and p = 0.000000011, respectively; of Hep −/− BDL vs. Flox BDL p = 0.000041, p = 0.013, and p = 0.0000031, respectively. For k , mRNA levels in HSCs, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham, p = 0.0000000000005, p = 0.000000067, and p = 0.0000000000 respectively; of Hep−/− BDL vs. Flox BDL p = 0.00000000050, p = 0.00025, and p = 0.0000000011 respectively. For l , mRNA levels in KCs, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham, p = 0.000041, p = 0.000010, and p = 0.00000035 respectively; of Hep−/− BDL vs. Flox BDL p = 0.84, p = 0.16, and p = 0.62 respectively. Statistical significance was determined by using two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: ALT alanine transaminase, AST aspartate aminotransferase, BDL bile duct ligation, Cho cholangiocytes, HSCs hepatic stellate cells, Hep hepatocytes, KCs Kupffer cells.

Journal: Nature Communications

Article Title: The role of forkhead box M1-methionine adenosyltransferase 2 A/2B axis in liver inflammation and fibrosis

doi: 10.1038/s41467-024-52527-8

Figure Lengend Snippet: a H&E, Sirius red, CK19, α-SMA, and F4/80 staining in Flox control and Foxm1 Hep−/− mice after BDL as compared to sham surgery. Liver fibrosis was measured by Sirius red staining ( n = 7 per group) ( b ) and hydroxyproline assay ( n = 5 per group) ( c ), liver injury by ALT ( n = 6 per group) ( d ) and AST ( n = 6 per group) ( e ) levels, macrophage number by F4/80 ( n = 6 per group) ( f ), and ductular proliferation by CK19 ( n = 4 per group) and myofibroblast differentiation by α-SMA staining ( n = 3 per group) ( g ). For b , Sirus red area/total area for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.00000000011 and p = 0.0000015, respectively. For c , hydroxyproline (ug/g, liver) for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.000058 and p = 0.0034, respectively. For d , ALT level (ug/L) for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.000000000014 and p = 0.00000000097, respectively. For e , AST level (ug/L) for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.0000000000001 and p = 0.00000025, respectively. For f , F4/80 positive number for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.0000000020 and p = 0.00080, respectively. For g , CK19/total area for Flox BDL vs. Sham and Hep−/− BDL vs. Flox BDL, p = 0.00000078 and p = 0.00026, respectively. Data are shown as mean ± SEM, ** p < 0.01, *** p < 0.001, **** p < 0.0001. h Protein expression of FOXM1, MATα2, and MAT2β in hepatocytes, cholangiocytes, HSCs, and KCs isolated from Flox control and Foxm1 Hep−/− mice ± BDL, Densitometry values for protein levels are summarized in Supplementary Fig. . mRNA levels of Foxm1 , Mat2a , and Mat2b in hepatocytes ( n = 6 animals in Flox + Sham, Foxm1 hep−/− + Sham, Flox + BDL groups and Foxm1 hep−/− + BDL group for foxm1 and mat2b mRNA; n = 4 in Flox + Sham, Foxm1 hep−/− + Sham, Flox + BDL groups and n = 6 in foxm1 hep−/− +BDL group for mat2a mRNA) ( i ), cholangiocytes ( n = 4 animals in Flox + Sham , foxm1 hep−/− + Sham, Flox + BDL groups and foxm1hep−/− + BDL groups for foxm1 mRNA; n = 5 in Flox + Sham and foxm1 hep−/− + Sham groups and n = 4 in Flox + BDL and Foxm1 hep-−/− + BDL groups for mat2a mRNA; n = 6 in Flox + Sham, Foxm1 hep−/− + Sham, Flox + BDL groups and foxm1 hep−/− + BDL group for mat2b mRNA) ( j ), HSCs ( n = 6 animals) ( k ), and KCs ( n = 4 animals in Flox + Sham, Flox + BDL groups, foxm1 hep−/− + BDL groups and n = 5 in foxm1 hep−/− + Sham for foxm1 mRNA; n = 5 in Flox + Sham and foxm1 hep−/− + Sham groups, Flox + BDL and Foxm1 hep−/− + BDL groups for mat2a mRNA; n = 5 in Flox + Sham, Flox + BDL and Foxm1 hep−/− + BDL groups and n = 6 in foxm1 hep−/− + Sham group for mat2b mRNA). l isolated from Flox control, Foxm1 Hep−/− mice ± BDL. Data are shown as mean fold of Flox control ± SEM, * p < 0.05, *** p < 0.001, **** p < 0.0001. p values obtained via two-tailed unpaired Student’s t tests. For i , mRNA levels in hepatocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham p = 0.0000045, p = 0.000030, and p = 0.000028 respectively; of Hep −/− BDL vs. Flox BDL p = 0.0000000099, p = 0.00024, p = 0.000099 respectively. For j , mRNA levels in cholangiocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham p = 0.00023, p = 0.00000026, and p = 0.000000011, respectively; of Hep −/− BDL vs. Flox BDL p = 0.000041, p = 0.013, and p = 0.0000031, respectively. For k , mRNA levels in HSCs, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham, p = 0.0000000000005, p = 0.000000067, and p = 0.0000000000 respectively; of Hep−/− BDL vs. Flox BDL p = 0.00000000050, p = 0.00025, and p = 0.0000000011 respectively. For l , mRNA levels in KCs, fold of Flox con of FOXM1, MAT2A, and MAT2B of Flox BDL vs. Sham, p = 0.000041, p = 0.000010, and p = 0.00000035 respectively; of Hep−/− BDL vs. Flox BDL p = 0.84, p = 0.16, and p = 0.62 respectively. Statistical significance was determined by using two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: ALT alanine transaminase, AST aspartate aminotransferase, BDL bile duct ligation, Cho cholangiocytes, HSCs hepatic stellate cells, Hep hepatocytes, KCs Kupffer cells.

Article Snippet: Human and mouse probes for MAT2A , MAT2B and FOXM1 , and the PCR Supermix were purchased from ThermoFisher.

Techniques: Staining, Control, Hydroxyproline Assay, Expressing, Isolation, Two Tailed Test, Ligation

a H&E, Sirius red, CK19, α-SMA, and F4/80 staining in Flox control and Foxm1 HSC−/− mice after BDL as compared to sham surgery. Liver fibrosis was measured by Sirius red staining ( n = 8 animals per group) ( b ) and hydroxyproline assay ( n = 6 animals per group) ( c ), liver injury by ALT ( n = 6 animals in Flox + Sham, Flox + BDL and Foxm1 HSC−/− + BDL groups and n = 5 in Foxm1 HSC−/− + Sham) ( d ) and AST ( n = 7 in Flox + Sham, Flox + BDL groups and n = 6 in Flox + BDL group and n = 5 in foxm1 HSC−/− + BDL) ( e ) levels, macrophage number by F4/80 ( n = 4 animals per group) ( f ), and ductular proliferation by CK19 staining ( n = 6 animals per group) and myofibroblast differentiation by α-SMA staining ( n = 3 animals per group) ( g ). For b , Sirius red area/total area of Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.0000000015 and p = 0.0000060 respectively. For c , hydroxyproline (ug/g, liver) for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.00000012 and p = 0.000094 respectively. For d , ALT level (ug/L) for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.000000041 and p = 0.0000013 respectively. For e , AST level (ug/L) for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.00000000066 and p = 0.00044 respectively. For f , F4/80 positive number for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.0000000071 and p = 0.0073 respectively. For g , Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL for CK19/total area, p = 0.000000000075 and p = 0.080 respectively; for α-SMA area/total area, p = 0.000026 and p = 0.00011 respectively. Data are shown as mean ± SEM, *** p < 0.001, **** p < 0.0001. h Protein expression of FOXM1, MATα2, and MAT2β in hepatocytes, cholangiocytes, HSCs, and KCs isolated from Flox control and Foxm1 HSC−/− mice ± BDL, Densitometry values for protein levels are summarized in Supplementary Fig. . mRNA levels of Foxm1 , Mat2a , and Mat2b in hepatocytes ( n = 6 per group) ( i ), cholangiocytes ( n = 6 per group) ( j ), HSCs ( n = 6 per group) ( k ), and KCs ( n = 6 per group) ( l ) isolated from Flox control and Foxm1 HSC−/− mice ± BDL. Data are shown as mean fold of Flox control ± SEM, **** p < 0.0001, and ns not significant. p values obtained via two-tailed unpaired Student’s t tests. For i , mRNA levels in Hepatocytes, fold of Flox con of FOXM1, MAT2A, MAT2B for Flox BDL vs. Sham, p = 0.0000000000004, p = 0.0000000000001 and p = 0.0000000000072 respectively; for HSC−/− BDL vs Flox BDL, p = 0.000000000022, p = 0.000000000020, and p = 0.000000054 respectively. For j , mRNA levels in Cholangiocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000000000000, p = 0.000000028, and p = 0.0000000085 respectively; for HSC−/− BDL vs Flox BDL, p = 0.17, p = 0.42, and p = 0.46 respectively. For k , mRNA levels in HSCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000052, p = 0.0000000000, and p = 0.0000000000 respectively; for HSC−/− BDL vs Flox BDL, p = 0.00000056, p = 0.00018, and p = 0.00000000002 respectively. For l , mRNA levels in KCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.000000000091, p = 0.00036, and p = 0.00000010 respectively; for HSC−/− BDL vs Flox BDL, p = 0.086, p = 0.61, and p = 0.15 respectively. Statistical significance was determined by using two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: ALT alanine transaminase, AST aspartate aminotransferase, BDL bile duct ligation, Cho cholangiocytes, HSCs hepatic stellate cells; Hep hepatocytes, KCs Kupffer cells.

Journal: Nature Communications

Article Title: The role of forkhead box M1-methionine adenosyltransferase 2 A/2B axis in liver inflammation and fibrosis

doi: 10.1038/s41467-024-52527-8

Figure Lengend Snippet: a H&E, Sirius red, CK19, α-SMA, and F4/80 staining in Flox control and Foxm1 HSC−/− mice after BDL as compared to sham surgery. Liver fibrosis was measured by Sirius red staining ( n = 8 animals per group) ( b ) and hydroxyproline assay ( n = 6 animals per group) ( c ), liver injury by ALT ( n = 6 animals in Flox + Sham, Flox + BDL and Foxm1 HSC−/− + BDL groups and n = 5 in Foxm1 HSC−/− + Sham) ( d ) and AST ( n = 7 in Flox + Sham, Flox + BDL groups and n = 6 in Flox + BDL group and n = 5 in foxm1 HSC−/− + BDL) ( e ) levels, macrophage number by F4/80 ( n = 4 animals per group) ( f ), and ductular proliferation by CK19 staining ( n = 6 animals per group) and myofibroblast differentiation by α-SMA staining ( n = 3 animals per group) ( g ). For b , Sirius red area/total area of Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.0000000015 and p = 0.0000060 respectively. For c , hydroxyproline (ug/g, liver) for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.00000012 and p = 0.000094 respectively. For d , ALT level (ug/L) for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.000000041 and p = 0.0000013 respectively. For e , AST level (ug/L) for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.00000000066 and p = 0.00044 respectively. For f , F4/80 positive number for Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL, p = 0.0000000071 and p = 0.0073 respectively. For g , Flox BDL vs. Sham and HSC−/− BDL vs. Flox BDL for CK19/total area, p = 0.000000000075 and p = 0.080 respectively; for α-SMA area/total area, p = 0.000026 and p = 0.00011 respectively. Data are shown as mean ± SEM, *** p < 0.001, **** p < 0.0001. h Protein expression of FOXM1, MATα2, and MAT2β in hepatocytes, cholangiocytes, HSCs, and KCs isolated from Flox control and Foxm1 HSC−/− mice ± BDL, Densitometry values for protein levels are summarized in Supplementary Fig. . mRNA levels of Foxm1 , Mat2a , and Mat2b in hepatocytes ( n = 6 per group) ( i ), cholangiocytes ( n = 6 per group) ( j ), HSCs ( n = 6 per group) ( k ), and KCs ( n = 6 per group) ( l ) isolated from Flox control and Foxm1 HSC−/− mice ± BDL. Data are shown as mean fold of Flox control ± SEM, **** p < 0.0001, and ns not significant. p values obtained via two-tailed unpaired Student’s t tests. For i , mRNA levels in Hepatocytes, fold of Flox con of FOXM1, MAT2A, MAT2B for Flox BDL vs. Sham, p = 0.0000000000004, p = 0.0000000000001 and p = 0.0000000000072 respectively; for HSC−/− BDL vs Flox BDL, p = 0.000000000022, p = 0.000000000020, and p = 0.000000054 respectively. For j , mRNA levels in Cholangiocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000000000000, p = 0.000000028, and p = 0.0000000085 respectively; for HSC−/− BDL vs Flox BDL, p = 0.17, p = 0.42, and p = 0.46 respectively. For k , mRNA levels in HSCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000052, p = 0.0000000000, and p = 0.0000000000 respectively; for HSC−/− BDL vs Flox BDL, p = 0.00000056, p = 0.00018, and p = 0.00000000002 respectively. For l , mRNA levels in KCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.000000000091, p = 0.00036, and p = 0.00000010 respectively; for HSC−/− BDL vs Flox BDL, p = 0.086, p = 0.61, and p = 0.15 respectively. Statistical significance was determined by using two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: ALT alanine transaminase, AST aspartate aminotransferase, BDL bile duct ligation, Cho cholangiocytes, HSCs hepatic stellate cells; Hep hepatocytes, KCs Kupffer cells.

Article Snippet: Human and mouse probes for MAT2A , MAT2B and FOXM1 , and the PCR Supermix were purchased from ThermoFisher.

Techniques: Staining, Control, Hydroxyproline Assay, Expressing, Isolation, Two Tailed Test, Ligation

a H&E, Sirius red, CK19, α-SMA, and F4/80 staining in Flox control and Foxm1 KC−/− mice after BDL as compared to sham surgery. Liver fibrosis was measured by Sirius red staining ( n = 8 animals per group), b and hydroxyproline assay ( n = 5 animals in Flox + Sham group and n = 6 in foxm1 KC−/− + Sham, Flox + BDL and foxm1 KC−/− + BDL groups) ( c ), liver injury by ALT ( n = 6 animals per group) ( d ) and AST ( n = 6 animals per group) ( e ) levels, macrophage number by F4/80 ( n = 6 animals per group) ( f ), and ductular proliferation by CK19 ( n = 4 animals per group) and myofibroblast differentiation by α-SMA staining ( n = 3 animals per group) ( g ). For b , Sirius red area/total area of Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.00000000049 and p = 0.00014 respectively. For c , hydroxyproline (ug/g, liver) for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.00030 and p = 0.012 respectively. For d , ALT level (ug/L) for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.0000000000000 and p = 0.000000000081 respectively. For e , AST level (ug/L) for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.0000000000001 and p = 0.00000025 respectively. For f , F4/80 positive number for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.0000000067 and p = 0.0000094 respectively. For g , Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL for CK19/total area, p = 0.00000010 and p = 0.039 respectively; for α-SMA area/total area, p = 0.000026 and p = 0.00033 respectively. Data are shown as mean ± SEM, * p < 0.05, *** p < 0.001, **** p < 0.0001. h Protein levels of FOXM1, MATα2, and MAT2β in hepatocytes, cholangiocytes, HSCs, and KCs isolated from Flox and Foxm1 KC−/− mice ± BDL, Densitometry values for protein levels are summarized in Supplementary Fig. . mRNA levels of Foxm1 , Mat2a , and Mat2b in hepatocytes ( n = 6 per group) ( i ), cholangiocytes ( n = 6 per group) ( j ), HSCs ( n = 6 per group) ( k ), and KCs ( n = 6 per group) ( l ) isolated from Flox control and Foxm1 KC−/− mice ± BDL. Data are shown as mean fold of Flox control ± SEM, *** p < 0.001, **** p < 0.0001, ns not significant. For i , mRNA levels in hepatocytes, fold of Flox con of FOXM1, MAT2A, MAT2B for Flox BDL vs. Sham, p = 0.000000012, p = 0.0000019 and p = 0.50 respectively; for KC −/− BDL vs Flox BDL, p = 0.00000086, p = 0.00060, and p = 0.0000000036 respectively. For j , mRNA levels in cholangiocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000000000001, p = 0.0000000000005, and p = 0.00000000046 respectively; for KC −/− BDL vs Flox BDL, p = 0.071, p = 0.064, and p = 0.061 respectively. For k , mRNA levels in HSCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000000086, p = 0.0000000002, and p = 0.000000000 respectively; for KC −/− BDL vs Flox BDL, p = 0.0000032, p = 0.0000095, and p = 0.0000078 respectively. For l , mRNA levels in KCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.000000040, p = 0.000000033, and p = 0.00000075 respectively; for KC −/− BDL vs Flox BDL, p = 0.00000000071, p = 0.00000097, and p = 0.0000059, respectively. Statistical significance was determined by using two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: ALT alanine transaminase, AST aspartate aminotransferase, BDL bile duct ligation, Cho cholangiocytes, HSCs hepatic stellate cells, Hep hepatocytes, KCs Kupffer cells.

Journal: Nature Communications

Article Title: The role of forkhead box M1-methionine adenosyltransferase 2 A/2B axis in liver inflammation and fibrosis

doi: 10.1038/s41467-024-52527-8

Figure Lengend Snippet: a H&E, Sirius red, CK19, α-SMA, and F4/80 staining in Flox control and Foxm1 KC−/− mice after BDL as compared to sham surgery. Liver fibrosis was measured by Sirius red staining ( n = 8 animals per group), b and hydroxyproline assay ( n = 5 animals in Flox + Sham group and n = 6 in foxm1 KC−/− + Sham, Flox + BDL and foxm1 KC−/− + BDL groups) ( c ), liver injury by ALT ( n = 6 animals per group) ( d ) and AST ( n = 6 animals per group) ( e ) levels, macrophage number by F4/80 ( n = 6 animals per group) ( f ), and ductular proliferation by CK19 ( n = 4 animals per group) and myofibroblast differentiation by α-SMA staining ( n = 3 animals per group) ( g ). For b , Sirius red area/total area of Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.00000000049 and p = 0.00014 respectively. For c , hydroxyproline (ug/g, liver) for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.00030 and p = 0.012 respectively. For d , ALT level (ug/L) for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.0000000000000 and p = 0.000000000081 respectively. For e , AST level (ug/L) for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.0000000000001 and p = 0.00000025 respectively. For f , F4/80 positive number for Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL, p = 0.0000000067 and p = 0.0000094 respectively. For g , Flox BDL vs. Sham and KC−/− BDL vs. Flox BDL for CK19/total area, p = 0.00000010 and p = 0.039 respectively; for α-SMA area/total area, p = 0.000026 and p = 0.00033 respectively. Data are shown as mean ± SEM, * p < 0.05, *** p < 0.001, **** p < 0.0001. h Protein levels of FOXM1, MATα2, and MAT2β in hepatocytes, cholangiocytes, HSCs, and KCs isolated from Flox and Foxm1 KC−/− mice ± BDL, Densitometry values for protein levels are summarized in Supplementary Fig. . mRNA levels of Foxm1 , Mat2a , and Mat2b in hepatocytes ( n = 6 per group) ( i ), cholangiocytes ( n = 6 per group) ( j ), HSCs ( n = 6 per group) ( k ), and KCs ( n = 6 per group) ( l ) isolated from Flox control and Foxm1 KC−/− mice ± BDL. Data are shown as mean fold of Flox control ± SEM, *** p < 0.001, **** p < 0.0001, ns not significant. For i , mRNA levels in hepatocytes, fold of Flox con of FOXM1, MAT2A, MAT2B for Flox BDL vs. Sham, p = 0.000000012, p = 0.0000019 and p = 0.50 respectively; for KC −/− BDL vs Flox BDL, p = 0.00000086, p = 0.00060, and p = 0.0000000036 respectively. For j , mRNA levels in cholangiocytes, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000000000001, p = 0.0000000000005, and p = 0.00000000046 respectively; for KC −/− BDL vs Flox BDL, p = 0.071, p = 0.064, and p = 0.061 respectively. For k , mRNA levels in HSCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.0000000086, p = 0.0000000002, and p = 0.000000000 respectively; for KC −/− BDL vs Flox BDL, p = 0.0000032, p = 0.0000095, and p = 0.0000078 respectively. For l , mRNA levels in KCs, fold of Flox con of FOXM1, MAT2A, and MAT2B for Flox BDL vs. Sham p = 0.000000040, p = 0.000000033, and p = 0.00000075 respectively; for KC −/− BDL vs Flox BDL, p = 0.00000000071, p = 0.00000097, and p = 0.0000059, respectively. Statistical significance was determined by using two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file. Abbreviations: ALT alanine transaminase, AST aspartate aminotransferase, BDL bile duct ligation, Cho cholangiocytes, HSCs hepatic stellate cells, Hep hepatocytes, KCs Kupffer cells.

Article Snippet: Human and mouse probes for MAT2A , MAT2B and FOXM1 , and the PCR Supermix were purchased from ThermoFisher.

Techniques: Staining, Control, Hydroxyproline Assay, Isolation, Two Tailed Test, Ligation

a Time courses of protein expression of FOXM1, MATα2, MAT2β, SMAD3, α–SMA, and COL1A1 after TGF-β1 treatment in LX-2 cells. b FOXM1, MATα2, MAT2β, SMAD3, α-SMA and COL1A1 protein levels after siRNA knockdown of FOXM1, MAT2A, or MAT2B in LX-2 cells after 24 h. c Protein levels of FOXM1, MATα2, MAT2β, SMAD3, α-SMA, and COL1A1 after FOXM1 overexpression with or without MAT2A or MAT2B siRNA knockdown and TGF–β1 treatment (20 ng/ml) for 24 h in LX-2 cells. d Effect of LPS on protein expression of FOXM1, MATα2, MAT2β, TNF-α, and IL-6 with or without siRNA knockdown of FOXM1, MAT2A or MAT2B in RAW 264.7 cells for 24 h. e Effects of LPS on protein expression of FOXM1, MATα2, MAT2β, TNF-α, and IL-6 with FOXM1 overexpression and MAT2A or MAT2B siRNA treatment for 24 h in RAW 264.7 cells (left panel) and KCs isolated from Flox control mice (right panel). Densitometry values for protein levels are summarized in Supplementary Fig. and Supplementary Fig. . n = 3 independent experiments. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: The role of forkhead box M1-methionine adenosyltransferase 2 A/2B axis in liver inflammation and fibrosis

doi: 10.1038/s41467-024-52527-8

Figure Lengend Snippet: a Time courses of protein expression of FOXM1, MATα2, MAT2β, SMAD3, α–SMA, and COL1A1 after TGF-β1 treatment in LX-2 cells. b FOXM1, MATα2, MAT2β, SMAD3, α-SMA and COL1A1 protein levels after siRNA knockdown of FOXM1, MAT2A, or MAT2B in LX-2 cells after 24 h. c Protein levels of FOXM1, MATα2, MAT2β, SMAD3, α-SMA, and COL1A1 after FOXM1 overexpression with or without MAT2A or MAT2B siRNA knockdown and TGF–β1 treatment (20 ng/ml) for 24 h in LX-2 cells. d Effect of LPS on protein expression of FOXM1, MATα2, MAT2β, TNF-α, and IL-6 with or without siRNA knockdown of FOXM1, MAT2A or MAT2B in RAW 264.7 cells for 24 h. e Effects of LPS on protein expression of FOXM1, MATα2, MAT2β, TNF-α, and IL-6 with FOXM1 overexpression and MAT2A or MAT2B siRNA treatment for 24 h in RAW 264.7 cells (left panel) and KCs isolated from Flox control mice (right panel). Densitometry values for protein levels are summarized in Supplementary Fig. and Supplementary Fig. . n = 3 independent experiments. Source data are provided as a Source Data file.

Article Snippet: Human and mouse probes for MAT2A , MAT2B and FOXM1 , and the PCR Supermix were purchased from ThermoFisher.

Techniques: Expressing, Knockdown, Over Expression, Isolation, Control

a Following BDL and culture of hepatocytes from flox mice and Foxm1 Hep−/− , EVs were extracted from the medium and used to treat HSCs from Foxm1 HSC−/− . b Following BDL and culture of HSCs from flox mice and Foxm1 HSC−/− , EVs were extracted from the medium and used to treat hepatocytes from Foxm1 Hep−/− . c , d Following BDL and culture of KCs from flox mice and Foxm1 KC−/− , EVs were extracted from the medium and used to treat ( c ) HSCs from Foxm1 HSC−/− or ( d ) hepatocytes from Foxm1 Hep−/− . e EVs extracted from hepatocytes of Flox and Foxm1 Hep −/− after BDL were used to treat KCs from Foxm1 KC −/− and ( f ) EVs extracted from HSCs of Flox and Foxm1 HSC −/− after BDL were used to treat KCs from Foxm1 KC−/− . All EV treatments were for 24 h after which protein expression of FOXM1, MATα2 and MAT2β were measured in cell lysates by western blotting. Densitometry values for protein levels are summarized in Supplementary Fig. , n = 3 independent experiments ( g ) Summary of key findings showing the FOXM1/MAT2A/MAT2B axis in the different liver cell types driving liver inflammation and fibrosis. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: The role of forkhead box M1-methionine adenosyltransferase 2 A/2B axis in liver inflammation and fibrosis

doi: 10.1038/s41467-024-52527-8

Figure Lengend Snippet: a Following BDL and culture of hepatocytes from flox mice and Foxm1 Hep−/− , EVs were extracted from the medium and used to treat HSCs from Foxm1 HSC−/− . b Following BDL and culture of HSCs from flox mice and Foxm1 HSC−/− , EVs were extracted from the medium and used to treat hepatocytes from Foxm1 Hep−/− . c , d Following BDL and culture of KCs from flox mice and Foxm1 KC−/− , EVs were extracted from the medium and used to treat ( c ) HSCs from Foxm1 HSC−/− or ( d ) hepatocytes from Foxm1 Hep−/− . e EVs extracted from hepatocytes of Flox and Foxm1 Hep −/− after BDL were used to treat KCs from Foxm1 KC −/− and ( f ) EVs extracted from HSCs of Flox and Foxm1 HSC −/− after BDL were used to treat KCs from Foxm1 KC−/− . All EV treatments were for 24 h after which protein expression of FOXM1, MATα2 and MAT2β were measured in cell lysates by western blotting. Densitometry values for protein levels are summarized in Supplementary Fig. , n = 3 independent experiments ( g ) Summary of key findings showing the FOXM1/MAT2A/MAT2B axis in the different liver cell types driving liver inflammation and fibrosis. Source data are provided as a Source Data file.

Article Snippet: Human and mouse probes for MAT2A , MAT2B and FOXM1 , and the PCR Supermix were purchased from ThermoFisher.

Techniques: Expressing, Western Blot

Journal: Journal of the American Chemical Society

Article Title: The Transition-State Structure for Human MAT2A from Isotope Effects

doi: 10.1021/jacs.7b05803

Figure Lengend Snippet: Summary of V/K Kinetic Isotope Effects, Intrinsic Kinetic Isotope Effects, Binding Isotope Effects and Calculated Kinetic Isotope Effects from QM Calculations for Human MAT2A at 5 Atomic Positions

Article Snippet: A synthetic plasmid encoding human MAT2A was designed in a modified pET vector (now deposited at AddGene).

Techniques: Binding Assay

Journal: Journal of the American Chemical Society

Article Title: The Transition-State Structure for Human MAT2A from Isotope Effects

doi: 10.1021/jacs.7b05803

Figure Lengend Snippet: MAT2A catalyzes the formation of SAM from ATP and methionine. KIE measurements used isotopically labeled substrates with the labels indicated. Each color corresponds to labels found on an individual substrate molecule.

Article Snippet: A synthetic plasmid encoding human MAT2A was designed in a modified pET vector (now deposited at AddGene).

Techniques: Labeling

Journal: Journal of the American Chemical Society

Article Title: The Transition-State Structure for Human MAT2A from Isotope Effects

doi: 10.1021/jacs.7b05803

Figure Lengend Snippet: (A) Geometric and electrostatic potential surface (EPS) maps for the MAT2A transition state. Red color designates partial negative charge while blue color designates partial positive charge. (B) Zoomed in EPS maps of the central sulfur atom for the MAT2A transition state (TS), S-adenosyl-L-methionine (SAM) and methionine (Met) are shown for comparison. The partial positive charge on the sulfonium group is significantly reduced in the transition state structure when compared to the SAM structure.

Article Snippet: A synthetic plasmid encoding human MAT2A was designed in a modified pET vector (now deposited at AddGene).

Techniques: Comparison

Proteins significantly regulated following RAPTA-T treatment determined from FITExP analysis of MCF-7 and MDA-MB-231 cells.

Journal: Scientific Reports

Article Title: Expression proteomics study to determine metallodrug targets and optimal drug combinations

doi: 10.1038/s41598-017-01643-1

Figure Lengend Snippet: Proteins significantly regulated following RAPTA-T treatment determined from FITExP analysis of MCF-7 and MDA-MB-231 cells.

Article Snippet: Human MAT2A plasmid was obtained from Addgene ( www.addgene.org , plasmid #53648) as a bacterial stab.

Techniques: Transmission Assay

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