rabbit polyclonal anti mat2a  (Proteintech)

Journal: The Journal of Experimental Medicine

Article Title: MYCT1–IFITM2/3 interaction links endothelial endolysosomal trafficking to white adipose tissue expansion

doi: 10.1084/jem.20251497

Figure Lengend Snippet: Key resources table

Article Snippet: Rabbit polyclonal anti-Mat2a , Proteintech , Cat No. 55309-1-AP, RRID:AB_2881303.

Techniques: Virus, Recombinant, Protease Inhibitor, Red Blood Cell Lysis, Control, In Situ, cDNA Synthesis, Bicinchoninic Acid Protein Assay, Isolation, Negative Control, Software

Journal: Nature Communications

Article Title: MAT2A promotes atherosclerotic plaque vulnerability by mediating epigenetic reprogramming of macrophages

doi: 10.1038/s41467-025-66121-z

Figure Lengend Snippet: A The enrollment of TCFA-positive patients defined by OCT and schematic representation of the patients screened in the discovery cohort. This figure was created using images from PowerPoint and BioRender. Created in BioRender. wan, p. (2025) https://BioRender.com/dmkgnf0 . B Partial least-squares discrimination analysis (PLS-DA) 3D score plots. C Volcano plot of significantly differential metabolites in monocytes. D Detection of monocytes methionine metabolism concentrations ( n = 38 subjects). E The final FCT was determined by averaging three measurements performed at the thinnest part of the fibrous cap. F Vulnerable plaque characteristics between patients with high and low SAM/SAH levels ( n = 38 subjects). G Multivariate logistic regression analysis revealing the relationship between high SAM/SAH levels and TCFA ( n = 76 for total subjects). H Representative OCT images of a normal coronary artery, a vulnerable plaque, and a stable plaque. Scale bar = 5 mm. I Schematic of the methionine metabolism pathway. J The mRNA levels in monocytes isolated from TCFA-positive and TCFA-negative individuals ( n = 38 subjects). UMAP visualization ( K ) and MAT2A expression ( L ) of human atherosclerotic carotid artery cells ( n = 15 patients, GSE253903 ). UMAP visualization ( M ) and MAT2A expression ( N ) of human atherosclerotic carotid artery myeloid cells. Representative immunofluorescence images ( O ) and quantification ( P ) of MAT2A and CD11b in carotid plaques from symptomatic and asymptomatic patients ( n = 6 subjects. Scale bar = 50 µm). C Two-tailed P-values were indicated. D The bar bands defined as 25th and 75th percentiles; the plot top line as median; two-tailed Mann–Whitney P -values are indicated. F The box shows the 25th, 50th and 75th percentiles of the data. The whiskers represent minimum and the maximum; Chi-squared test and two-tailed Mann–Whitney P -values was used. G The error bars represent the 95% CI for the OR, with their centers (red squares) indicating the point estimates; two-tailed P -values are shown. ( J , P ) Data are presented as the mean ± SD; two-tailed P -values are shown. L , N Log normalized using the ‘NormalizeData’ function in Seurat with default parameters. CKD chronic kidney disease, FCT fibrous cap thickness, MAT2A methionine adenosyltransferase Ⅱ alpha, OCT optical coherence tomography, SAH, S-adenosyl-homocysteine, SAM S-adenosyl-methionine, TCFA thin-cap fibroatheroma, UMAP uniform manifold approximation and projection. Source data are provided as a Source Data file.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against MAT2A (1:200, sc-398917, Santa Cruz Biotechnology, USA), CD11b (1:200, ab128797, Abcam, UK), CD68 (1:100, ab125212, Abcam, UK), and H3K4me3 (1:500, 39060, Active Motif, Shanghai, China).

Techniques: Isolation, Expressing, Immunofluorescence, Two Tailed Test, MANN-WHITNEY, Tomography

Journal: Nature Communications

Article Title: MAT2A promotes atherosclerotic plaque vulnerability by mediating epigenetic reprogramming of macrophages

doi: 10.1038/s41467-025-66121-z

Figure Lengend Snippet: Both groups of 8-week-old female and male MAT2A fl/fl ApoE -/- and MAT2A CKO ApoE -/- mice were fed an HFD for 16 weeks. A Schematic figure showing the experimental strategy for the HFD feeding and the subsequent analysis. This figure was created using images from Photoshop and BioRender. Created in BioRender. wan, p. (2025) https://BioRender.com/3iunh0m . B SAM and SAH concentrations as well as SAM/SAH ratio for mice monocyte cell lysate mass spectrometry samples ( n = 6). C Representative photographs of atherosclerotic plaques in the aortic arches and their branches in the 2 groups ( n = 10 mice per genotype). Representative images and quantification of Oil Red O-stained aortas ( D , n = 10 samples. Scale bar = 2 mm) and aortic roots ( E , n = 10 independent experiments. Scale bar = 200 µm). F , H , E and Masson’s trichrome staining of representative aortic root sections. Scale bar = 200 µm. G Quantification of plaque area, necrotic core, and collagen ( n = 10 samples). H and I , Immunohistochemistry of CD68 + macrophages and vulnerability index in the aortic roots ( n = 10 samples). Scale bar = 100 µm. Data are presented as the mean ± SD and comparisons were made using unpaired Student’s t -test; the two-tailed P -values are shown. α-SMA α-smooth muscle actin, HFD high-fat diet. Source data are provided as a Source Data file.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against MAT2A (1:200, sc-398917, Santa Cruz Biotechnology, USA), CD11b (1:200, ab128797, Abcam, UK), CD68 (1:100, ab125212, Abcam, UK), and H3K4me3 (1:500, 39060, Active Motif, Shanghai, China).

Techniques: Mass Spectrometry, Staining, Immunohistochemistry, Two Tailed Test

Journal: Nature Communications

Article Title: MAT2A promotes atherosclerotic plaque vulnerability by mediating epigenetic reprogramming of macrophages

doi: 10.1038/s41467-025-66121-z

Figure Lengend Snippet: A Bubble chart of GO term enrichment analysis of downregulated DEGs based on RNA-seq in BMDMs of MAT2A CKO compared with MAT2A fl/fl mice. Flow cytometry was performed to quantify CD45 + CD11b + CD115 + Ly6C hi monocytes from bone marrow or peripheral blood ( B , n = 6 independent experiments), MHC Ⅱ and CCR2 ( C , n = 6 samples) on CD45 + CD68 + macrophages from aortic plaques of MAT2A CKO ApoE -/- and MAT2A fl/fl ApoE -/- mice fed a 16-week HFD. D – J Analysis of BMDMs from MAT2A CKO and MAT2A fl/fl mice treated with or without SAM (200 µM) following LPS and IFN-γ stimulation. Representative flow cytometry plots and quantification of CD45 + CD11b + CD86 + macrophages ( D , n = 6 samples), transwell ( E , n = 6 samples, Scale bar = 100 µm), and wound assay ( F , n = 6 samples. Scale bar = 500 µm). The mRNA levels of Il1b , Il6 , Nos2 ( G , n = 6 samples) and Ccl2 , Ccl7 and Cxcl1 ( H , n = 6 samples) measured by qRT-PCR. I , J The THP-1 cells were transfected with siNC or siMAT2A followed by LPS and IFN-γ treatment with or without SAM (200 µM). The mRNA levels of Il1b , Il6 , Nos2, Ccl2 , Ccl7 and Cxcl1 were analyzed by qRT-PCR ( n = 6 samples). A P -values were derived from hypergeometric distribution. Data are presented as the mean ± SD. B , C Unpaired Student’s t -test was used; two-tailed P values are shown. D – J One-way ANOVA was used; the adjusted P -values are shown. 8-week-old female and male mice were both used. BL blood, BM bone marrow, BMDMs bone marrow-derived macrophages, DEGs differentially expressed genes, GO gene ontology, IFN-γ interferon-γ, LPS lipopolysaccharide. Source data are provided as a Source Data file.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against MAT2A (1:200, sc-398917, Santa Cruz Biotechnology, USA), CD11b (1:200, ab128797, Abcam, UK), CD68 (1:100, ab125212, Abcam, UK), and H3K4me3 (1:500, 39060, Active Motif, Shanghai, China).

Techniques: RNA Sequencing, Flow Cytometry, Quantitative RT-PCR, Transfection, Derivative Assay, Two Tailed Test

Journal: Nature Communications

Article Title: MAT2A promotes atherosclerotic plaque vulnerability by mediating epigenetic reprogramming of macrophages

doi: 10.1038/s41467-025-66121-z

Figure Lengend Snippet: A Immunoblots of H3K4me3, H3K36me3, and H3K27me3 levels in BMDMs from MAT2A fl/fl and MAT2A CKO mice ( n = 3). B Representative immunofluorescence images and quantification for H3K4me3 and CD68 in aortic root plaque from MAT2A fl/fl ApoE -/- and MAT2A CKO ApoE -/- mice fed a 16-week HFD ( n = 7). Scale bar = 100 µm. C – G Analysis of BMDMs between MAT2A CKO and MAT2A fl/fl mice. C Heatmaps for H3K4me3 binding peaks in BMDMs from MAT2A fl/fl and MAT2A CKO mice. D Venn diagram reflecting overlapping downregulated genes with H3K4me3 modification combining the CUT&Tag and RNA-seq data. Bubble charts of GO ( E ) and KEGG ( F ) analysis according to downregulated genes with decreased H3K4me3 modification. G IGV tracks revealing the results of CUT&Tag reads (H3K4me3 binding) distributions in indicated genes. H , I Analysis of BMDMs between MAT2A CKO and MAT2A fl/fl mice treated with or without SAM (200 µM) in the presence of LPS and IFN-γ. H ChIP-qPCR validation of H3K4me3 enrichment on the promoter regions of Aim2 , Ccl2 , and Mmp9 ( n = 6 samples). I ELISA analysis of AIM2, CCL2, and MMP9 in cellular supernatant ( n = 6 independent experiments). Data are presented as the mean ± SD. A , B Unpaired Student’s t -test was used; two-tailed P values are shown. E P -values were derived from hypergeometric distribution. H and I One-way ANOVA was used; the adjusted P -values are shown. Both 8-week-old female and male mice were used. ChIP-qPCR chromatin immunoprecipitation-qPCR, H3K4me3 histone h3 lysine 4 trimethylation, H3K27me3 histone H3 lysine 27 trimethylation, H3K36me3 histone h3 lysine 36 trimethylation, KEGG kyoto encyclopedia of genes and genomes, TSS translation start site. Source data are provided as a Source Data file.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against MAT2A (1:200, sc-398917, Santa Cruz Biotechnology, USA), CD11b (1:200, ab128797, Abcam, UK), CD68 (1:100, ab125212, Abcam, UK), and H3K4me3 (1:500, 39060, Active Motif, Shanghai, China).

Techniques: Western Blot, Immunofluorescence, Binding Assay, Modification, RNA Sequencing, ChIP-qPCR, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Derivative Assay, Chromatin Immunoprecipitation

Journal: Nature Communications

Article Title: MAT2A promotes atherosclerotic plaque vulnerability by mediating epigenetic reprogramming of macrophages

doi: 10.1038/s41467-025-66121-z

Figure Lengend Snippet: A – I The HFD-fed ApoE -/- mice were received either FIDAS-5 or vehicle every other day for 6 weeks. A, Flowchart illustrating the experimental procedure. This figure was created using images from Photoshop and BioRender. Created in BioRender. wan, p. (2025) https://BioRender.com/h76jjnw . B Representative photographs of atherosclerotic plaques in the aortic arches and their branches. Representative images and quantification of Oil Red O-stained aortas ( C , n = 8. Scale bar = 2 mm) and aortic roots ( D and E , n = 8. Scale bar = 200 µm). F Representative images of H&E and Masson’s trichrome-stained aortic roots. Scale bar = 200 µm. G Quantification of plaque area, percentage of necrotic core, and collagen of aortic roots ( n = 8 per group). H , I Immunohistochemistry of CD68 + macrophages and vulnerability index in the aortic root plaque of FIDAS-5 treated mice, n = 8 per group. Scale bar = 100 µm. J – R Representative images of female and male mice fed MRD or CD for 6 weeks after a 10-week HFD. J Schematic figure showing the experimental strategy and subsequent analysis. This figure was created using images from Photoshop and BioRender. Created in BioRender. wan, p. (2025) https://BioRender.com/h76jjnw . K Representative images of aortic arches and their branches in the indicated group. Representative images and quantification of Oil Red O-stained aortas ( L , n = 8. Scale bar = 2 mm) and aortic roots ( M and N , n = 8. Scale bar = 200 µm). O Representative images of H&E and Masson’s trichrome-stained aortic root. P Quantification of plaque area, percentage of necrotic core, and collagen, n = 8. Scale bar = 200 µm. Q and R Immunohistochemical staining CD68 + macrophages and vulnerability index, n = 8. Scale bar = 100 µm. Data are presented as mean ± SD and comparisons were made using unpaired Student’s t -test; two-tailed P values are shown. 8-week-old female and male ApoE -/- mice were used. FIDAS-5, a MAT2A inhibitor; MRD, methionine-restricted diet. Source data are provided as a Source Data file.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against MAT2A (1:200, sc-398917, Santa Cruz Biotechnology, USA), CD11b (1:200, ab128797, Abcam, UK), CD68 (1:100, ab125212, Abcam, UK), and H3K4me3 (1:500, 39060, Active Motif, Shanghai, China).

Techniques: Staining, Immunohistochemistry, Immunohistochemical staining, Two Tailed Test

Journal: Nature Communications

Article Title: MAT2A promotes atherosclerotic plaque vulnerability by mediating epigenetic reprogramming of macrophages

doi: 10.1038/s41467-025-66121-z

Figure Lengend Snippet: A Norepinephrine levels in serum from TCFA-positive and TCFA-negative individuals ( n = 38 independent experiments). B Correlation between serum the ratio of SAM/SAH and norepinephrine levels. C The percentage of patients presenting with TCFA in 3 coronary arteries varied according to their levels of the SAM/SAH ratio and norepinephrine. D MAT2A expression in peripheral blood monocytes from 6-OHDA-treated ApoE -/- mice ( n = 3, one representative experiment out of three was shown). E Representative images and quantification of H&E and Oil Red O staining in aortic root treated with 6-OHDA (250 mg/kg) and AAV-MAT2A, n = 6. Scale bar = 100 µm. F Immunoblots of aortic plaques from ApoE -/- mice treated with 6-OHDA ( n = 3). G , Mat2a mRNA expression in BMDMs pre-treated with 20 nM RAPA followed by 10 mM norepinephrine ( n = 6, one representative experiment out of three was shown). H Transcript levels of c-Myc and Mat2a in BMDMs transfected with the indicated siRNAs ( n = 6). I Occupancy analysis of c-MYC by ChIP-qPCR in BMDMs treated with DMSO vehicle or RAPA (20 nM) ( n = 6). A Two-tailed Mann–Whitney P -values are indicated; Data are shown as median with IQR. B Pearson correlation coefficient test was used; the regression coefficients and two-tailed P -values are shown. C Chi-squared test was used; Data are shown as %. D – F Unpaired Student’s t -test was used; the two-tailed P -values are shown; Data are presented as mean ± SD. G , I Two-way ANOVA was used; The adjusted P -values are shown; Data are presented as mean ± SD. H One-way ANOVA was used; The adjusted P -values are shown. 8-week-old female and male ApoE -/- mice were used. Data are presented as mean ± SD. AAV adeno-associated virus, RAPA rapamycin, 6-OHDA 6-Hydroxydopamine. Source data are provided as a Source Data file.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against MAT2A (1:200, sc-398917, Santa Cruz Biotechnology, USA), CD11b (1:200, ab128797, Abcam, UK), CD68 (1:100, ab125212, Abcam, UK), and H3K4me3 (1:500, 39060, Active Motif, Shanghai, China).

Techniques: Expressing, Staining, Western Blot, Transfection, ChIP-qPCR, Two Tailed Test, MANN-WHITNEY, Virus

Journal: Nature Communications

Article Title: MAT2A promotes atherosclerotic plaque vulnerability by mediating epigenetic reprogramming of macrophages

doi: 10.1038/s41467-025-66121-z

Figure Lengend Snippet: A Study design in the validation cohort. This figure was created using images from PowerPoint and BioRender. Created in BioRender. wan, p. (2025) https://BioRender.com/qbkjhvs . Detection of monocytes SAM and SAH concentrations ( B ), and SAM/SAH ratio ( C ) of TCFA-positive and TCFA-negative individuals ( n = 100 subjects). D Serum norepinephrine levels from TCFA-positive and TCFA-negative individuals ( n = 100 independent experiments). Qualitative and quantitative OCT analysis of vulnerable plaque characteristics between patients with high and low SAM/SAH levels ( E ) or with high and low norepinephrine levels ( F ) ( n = 100 independent experiments). Pearson correlation analysis of monocytes SAM/SAH ratio ( G ) or serum norepinephrine levels ( H ) with the thinnest FCT and mean lipid arc. I Two-tailed Pearson’s linear regression analysis between the SAM/SAH ratio and norepinephrine levels. J Multivariate logistic regression analysis depicting the relationship between the SAM/SAH ratio, norepinephrine levels, and TCFA ( n = 200 for total subjects). K Receiver operating characteristic curve of SAM/SAH, norepinephrine, and combined both for TCFA. L HR for incident 5-year MACE based on multivariable Cox proportional hazards regression analysis. Adjusted for age, gender, traditional coronary risk factors, and statin at discharge ( n = 200 for total subjects). M A Kaplan-Meier survival curve plots the 5-year MACE-free survival among 4 subgroups. N MAT2A-mediated monocyte methionine metabolism is closely associated with the presence of TCFA in patients. MAT2A, which is induced by the norepinephrine–mTOR–c-MYC axis, is critical for endowing monocytes/macrophages with proinflammatory state and migratory capacity during the development of atherosclerosis through H3K4me3 modification. Myeloid-specific MAT2A ablation, pharmacological blockade with FIDAS-5, or a low-methionine diet attenuate monocyte/macrophage inflammation and migration, thereby reducing atherosclerotic progression and plaque vulnerability. This figure was created using images from PowerPoint and BioRender. Created in BioRender. wan, p. (2025) https://BioRender.com/gylosq5 . B – D Two-tailed Mann–Whitney P -values are indicated. E , F The box shows the 25th, 50th and 75th percentiles of the data. The whiskers represent minimum and the maximum. Data are presented as % or median with IQR, Chi-squared test or Mann-Whitney test was used, and two-tailed P -values were calculated. G – I Pearson correlation coefficient test was used; the regression coefficients and two-tailed P -values are shown. J Data points indicate OR, and 95% confidence intervals are represented by line length; two-tailed P -values are shown. K The 95% confidence interval is shown between brackets. L Data points indicate HR and 95% CI were represented by line length. M P -values were calculated with log rank test. HR hazard ratio. Source data are provided as a Source Data file.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against MAT2A (1:200, sc-398917, Santa Cruz Biotechnology, USA), CD11b (1:200, ab128797, Abcam, UK), CD68 (1:100, ab125212, Abcam, UK), and H3K4me3 (1:500, 39060, Active Motif, Shanghai, China).

Techniques: Biomarker Discovery, Two Tailed Test, Modification, Migration, MANN-WHITNEY

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A novel role of secreted methionine adenosyltransferase α2 in colorectal liver metastases

doi: 10.1186/s13046-025-03599-x

Figure Lengend Snippet: Phosphorylation of RTKs in RKO cells treated with MATα2-t

Article Snippet: The slides were deparaffinized, hydrated, and stained for MATα1 (cat. H0004143-M01; Abnova, Taipei City, Taiwan) MATα2 (cat. NB110-94158; Novus, St. Charles, MI), and hepatocyte-specific antigen using extended antigen retrieval (antigen unmasking solution, cat. H-3301-250; Vector Laboratories, Burlingame, CA) as we described [ ].

Techniques: Phospho-proteomics

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A novel role of secreted methionine adenosyltransferase α2 in colorectal liver metastases

doi: 10.1186/s13046-025-03599-x

Figure Lengend Snippet: MATα2 is secreted by CRC cells in extracellular vesicles. A Exopred and Exocarta software predicts MATα2 is secreted via exosomes ( B ) CRC cells transfected to overexpress DDK-MAT2A (M2A) secreted more MATα2 when compared to empty vector (EV). C NanoSight analysis of extracellular vesicles isolated from culture media of CRC cells transfected to overexpress MAT2A (MAT2A OE) and empty vector (EVec) with peak corresponding to size range of exosomes. D Images from NanoSight analysis of extracellular vesicles. E Immunofluorescence microscopy of human hepatocytes treated with exosomes isolated from culture media from CRC transfected to overexpress DDK-MAT2A and empty vector (EVec) shows internalization of EV-MATα2 with localizing to the nuclues via DAPI staining

Article Snippet: The slides were deparaffinized, hydrated, and stained for MATα1 (cat. H0004143-M01; Abnova, Taipei City, Taiwan) MATα2 (cat. NB110-94158; Novus, St. Charles, MI), and hepatocyte-specific antigen using extended antigen retrieval (antigen unmasking solution, cat. H-3301-250; Vector Laboratories, Burlingame, CA) as we described [ ].

Techniques: Software, Transfection, Plasmid Preparation, Isolation, Immunofluorescence, Microscopy, Staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A novel role of secreted methionine adenosyltransferase α2 in colorectal liver metastases

doi: 10.1186/s13046-025-03599-x

Figure Lengend Snippet: EV-MATα2 is internalized by hepatocytes and alters MAT1A and MAT2A expression A mRNA levels of MAT1A and MAT2A isolated from human hepatocytes treated with exosomes from CRC cells. B Western blot analysis of endogenous nuclear and cytosolic MATα2 in human hepatocytes treated with exosomes from CRC cells. Lamin B1 and tubulin were used as loading controls for nuclear and cytoplasmic fractions, respectively. Mean ± SEM from n = 3, * p < 0.05 vs. EVec exo. C-D Confocal microscopy of human liver spheroids treated with exosomes from CRC cells transfected with empty vector (EVec) or MAT2A-His vector at day 7 for 24 h showing effect on MATα1 and MATα2-His expression. E mRNA levels of MAT1A and MAT2A isolated from human liver spheroids after the exosome treatment. Mean ± SEM from n = 3, * p < 0.02 vs.control and † p < 0.05 vs. EVec exo

Article Snippet: The slides were deparaffinized, hydrated, and stained for MATα1 (cat. H0004143-M01; Abnova, Taipei City, Taiwan) MATα2 (cat. NB110-94158; Novus, St. Charles, MI), and hepatocyte-specific antigen using extended antigen retrieval (antigen unmasking solution, cat. H-3301-250; Vector Laboratories, Burlingame, CA) as we described [ ].

Techniques: Expressing, Isolation, Western Blot, Confocal Microscopy, Transfection, Plasmid Preparation, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A novel role of secreted methionine adenosyltransferase α2 in colorectal liver metastases

doi: 10.1186/s13046-025-03599-x

Figure Lengend Snippet: Integrated analysis of MATα2 genomic binding profiles in CRC cells. A Heatmap showing the read density distribution of MATα2 ChIP-seq peaks across all human chromosomes. Read intensities are color-coded from low (purple) to high (yellow) density. B Genomic annotation of MATα2 binding peaks. C Top five DNA-binding motifs enriched within MATα2-bound peaks ranked by motif enrichment score. D Pathway enrichment analysis of MATα2-associated genes. E Predicted consensus sequences logos representing the top three de novo motifs identified from MATα2 binding sites by Jaspar software. F ChIP-seq tracks showing MATα2 (blue), RNA polymerase II (POL II; red) and input control (black) signals across the MAT1A (lower) or MAT2A (upper) locus. The Y-axis represents normalized read enrichment. The bottom track shows the gene structure with exons (black boxes) and introns (dashed lines)

Article Snippet: The slides were deparaffinized, hydrated, and stained for MATα1 (cat. H0004143-M01; Abnova, Taipei City, Taiwan) MATα2 (cat. NB110-94158; Novus, St. Charles, MI), and hepatocyte-specific antigen using extended antigen retrieval (antigen unmasking solution, cat. H-3301-250; Vector Laboratories, Burlingame, CA) as we described [ ].

Techniques: Binding Assay, ChIP-sequencing, Software, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A novel role of secreted methionine adenosyltransferase α2 in colorectal liver metastases

doi: 10.1186/s13046-025-03599-x

Figure Lengend Snippet: EV-MATα2 acts as a transcription factor to alter MAT1A and MAT2A expression. A Promoter activities in human hepatocytes transfected with human MAT1A or MAT2A promoter constructs and then treated with exosomes from RKO cells expressing empty vector (EVec exo) or MAT2A (EV-MATα2) as described in Methods. B ChIP analysis of the human MAT1A and ( C ) human MAT2A promoters showing binding of MATα2-His to different regions of the promoters. Mean ± SEM from n = 8, * p < 0.05 and ** p < 0.01 vs. EVec exo for MAT1A promoter; n = 7, * p < 0.04 vs. EVec exo for MAT2A promoter

Article Snippet: The slides were deparaffinized, hydrated, and stained for MATα1 (cat. H0004143-M01; Abnova, Taipei City, Taiwan) MATα2 (cat. NB110-94158; Novus, St. Charles, MI), and hepatocyte-specific antigen using extended antigen retrieval (antigen unmasking solution, cat. H-3301-250; Vector Laboratories, Burlingame, CA) as we described [ ].

Techniques: Expressing, Transfection, Construct, Plasmid Preparation, Binding Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A novel role of secreted methionine adenosyltransferase α2 in colorectal liver metastases

doi: 10.1186/s13046-025-03599-x

Figure Lengend Snippet: EV-MATα2 induces MAT2A expression and oncogenic activity in RKO cells. A RKO cells were treated with exosomes from RKO cells expressing empty vector (EVec exo) or MAT2A-His-Tag vector (EV-MATα2) as described in Methods and cell entry was visualized under fluorescent microscopy using His-tag antibody. B Real-time PCR shows the effect at the MAT2A mRNA level. Mean ± SEM from n = 3, * p < 0.02 vs. EVec exo. C Western blotting was done in total cell lysate, cytoplasmic and nuclear fractions showing increased MATα2 levels. Densitometry were measured by ImageJ. Mean ± SEM from n = 3, * p < 0.002 for total lysate, * p < 0.02 for cytoplasmic, * p < 0.03 for nuclear fractions vs. EVec exo. D ChIP analysis of the human MAT2A promoter showing binding of MATα2-His to different predicted motifs. Mean ± SEM from n = 3, * p < 0.05 vs. EVec exo. Effects of the same treatments on EdU ( E ) ( n = 3, * p < 0.05 and ** p < 0.01 vs. control), migration ( F ) ( n = 3, * p < 0.004 and ** p < 0.0001 vs. 0 h EVec exo, † p < 0.003 vs. 24 h EV-MATα2), and invasion ( G ) ( n = 3, * p < 0.008 vs. EVec exo)

Article Snippet: The slides were deparaffinized, hydrated, and stained for MATα1 (cat. H0004143-M01; Abnova, Taipei City, Taiwan) MATα2 (cat. NB110-94158; Novus, St. Charles, MI), and hepatocyte-specific antigen using extended antigen retrieval (antigen unmasking solution, cat. H-3301-250; Vector Laboratories, Burlingame, CA) as we described [ ].

Techniques: Expressing, Activity Assay, Plasmid Preparation, Microscopy, Real-time Polymerase Chain Reaction, Western Blot, Binding Assay, Control, Migration

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A novel role of secreted methionine adenosyltransferase α2 in colorectal liver metastases

doi: 10.1186/s13046-025-03599-x

Figure Lengend Snippet: MAT expression in adjacent hepatocytes is altered in human CRLM and patients with CRC secrete two forms of MATα2. A , B Human tissue microarray including normal human liver (NHL, n = 3) and CRLM ( n = 32) were examined using IHC (x100) for MATα1, MATα2, and hepatocyte specific antigen (HSA). Boxed areas are magnified, MATα1 and MATα2 staining in hepatocytes was analyzed by Image J and summarized in the graphs. Mean ± SEM, * p < 0.005 for MATα1, * p < 0.04 for MATα2 vs. NHL. C Human plasma from 5 healthy controls (HC) and 10 CRC patients were western blotted for MATα2 to detect full-length MATα2 (MATα2-fl) and truncated MATα2 (MATα2-t), with RKO lysate as input control. Mean ± SEM. * p < 0.05 vs. HC, ** p < 0.001 vs. HC

Article Snippet: The slides were deparaffinized, hydrated, and stained for MATα1 (cat. H0004143-M01; Abnova, Taipei City, Taiwan) MATα2 (cat. NB110-94158; Novus, St. Charles, MI), and hepatocyte-specific antigen using extended antigen retrieval (antigen unmasking solution, cat. H-3301-250; Vector Laboratories, Burlingame, CA) as we described [ ].

Techniques: Expressing, Microarray, Staining, Clinical Proteomics, Western Blot, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A novel role of secreted methionine adenosyltransferase α2 in colorectal liver metastases

doi: 10.1186/s13046-025-03599-x

Figure Lengend Snippet: Cancer cells secrete more truncated MATα2, which is required for survival. A Medium from RKO cells overexpressing MAT2A-His or empty vector (EVec) was separated into exosomes and EV-free media that only has truncated MATα2 (MATα2-t). Note MATα2-His has higher MW than full length endogenous MATα2, which has the same MW as MATα2-t-His. Endogenous MATα2-t has the lowest MW. B MATα2 protein sequence and predicted cleavage site based on PrediSI is at proline 30. C RKO (CRC), MiaPACA (pancreatic adenocarcinoma) and RV1 (prostate adenocarcinoma) cells secrete more MATα2-t as compared to the respective non-malignant cells (HCoEpC, HPDE, RWPE1). Mean ± SEM from n = 3, * p < 0.01 vs. HCoEpC cells; * p < 0.03 and † p < 0.01 vs. HPDE cells; * p < 0.03 and † p < 0.001 vs. RWPE1 cells. D RKO and HT29 cells treated with anti-MATα2 (20 µg/ml) for 48 h and TUNEL staining shows CRC cells underwent apoptosis. E MTT assay in RKO and HT29 cells treated with anti-MATα2 shows a fall in viability. Mean ± SEM from n = 3, * p < 0.03 and † p < 0.04 vs. control. F RKO cells were transfected with MATα2-DDK for 48 h and increasing amount of anti- MATα2 Ab was added, followed by pull-down of MATα2 Ab using beads, then western blotted the pull-down with anti-DDK Ab. The MATα2 antibody was able to bring down freely secreted MATα2 in a dose-dependent manner. This correlated with a dose-dependent increase in active caspase 3

Article Snippet: The slides were deparaffinized, hydrated, and stained for MATα1 (cat. H0004143-M01; Abnova, Taipei City, Taiwan) MATα2 (cat. NB110-94158; Novus, St. Charles, MI), and hepatocyte-specific antigen using extended antigen retrieval (antigen unmasking solution, cat. H-3301-250; Vector Laboratories, Burlingame, CA) as we described [ ].

Techniques: Plasmid Preparation, Sequencing, TUNEL Assay, Staining, MTT Assay, Control, Transfection, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A novel role of secreted methionine adenosyltransferase α2 in colorectal liver metastases

doi: 10.1186/s13046-025-03599-x

Figure Lengend Snippet: Secreted MATα2-t activates FAK and is required to maintain MAT2A expression. A RKO cells were treated with EV-free media containing MATα2-t as described in Methods and western blotted for pFAK and total FAK. Mean ± SEM from n = 3, * p < 0.04 vs. EVec ( B ) RKO cells were treated with anti-MATα2 Ab for 48 h and western blotted for pFAK, total FAK, pro-caspase 3 and active caspase 3. Mean ± SEM from n = 3, * p < 0.03 vs. control. C FAK and MAT2A mRNA levels in RKO cells from the above treatments were measured by real-time PCR. Mean ± SEM from n = 3, * p < 0.002 vs. EVec; * p < 0.002 vs. control. D RKO cells were CRISPR/Cas9 gene edited (HDR) to mutate proline to leucine at position 30 (canonical motif: PDLD) and at positions 131 and 133 glycine to leucine (non-canonical motif: GXGD); western blotted for pFAK and FAK. Mean ± SEM from n = 5–6, * p < 0.01 vs. wild-type (WT) for PDLD. E Immunoblotting of secreated MATα2 (MATα2-t) in culture media from RKO cells gene edited PDLD motif. Mean ± SEM from n = 3, * p < 0.04 vs. WT. F PDLD gene edited RKO and HT29 cells exhibited increased apoptosis on TUNEL staining (blue for RKO and brown for HT29 depending to pH culture media)

Article Snippet: The slides were deparaffinized, hydrated, and stained for MATα1 (cat. H0004143-M01; Abnova, Taipei City, Taiwan) MATα2 (cat. NB110-94158; Novus, St. Charles, MI), and hepatocyte-specific antigen using extended antigen retrieval (antigen unmasking solution, cat. H-3301-250; Vector Laboratories, Burlingame, CA) as we described [ ].

Techniques: Expressing, Western Blot, Control, Real-time Polymerase Chain Reaction, CRISPR, TUNEL Assay, Staining

Journal: Cell reports

Article Title: IGF2BP3 redirects glycolytic flux to promote one-carbon metabolism and RNA methylation

doi: 10.1016/j.celrep.2025.116330

Figure Lengend Snippet: (A) Western blot analysis of histone methylation (H3K4me1 and H3K4me4) in SEM and Lin− MLL-Af4 cells, control or depleted for IGF2BP3; n = 3. (B) Dot blot analysis of m 6 A modification (left) and methylene blue staining in SEM cells, control or depleted for IGF2BP3. (C) ELISA measurement of m 6 A modification on RNA isolated from SEM, Lin− MLL-Af4, and NALM6 cells ( n = 4 for SEM and Lin− MLL-Af4, n = 5 [sg5 = 3] for NALM6). (D) Bar plot (left) and pie chart (right) depicting the m 6 A peak distribution across genomic locations from the m 6 A-eCLIP data in SEM control and IGF2BP3-depleted cells. (E) Metagene plots depicting the changes in the m 6 A peak coverage across the transcriptome in SEM control and IGF2BP3-depleted cells. (F) Volcano plot (top) for genes showing differential m 6 A RNA methylation after IGF2BP3 depletion and IGF2BP3 targets defined by eCLIP analysis. Gray dashed lines indicate the significant cutoffs for differential expression (±1) and the adjusted p value (0.05). Hypomethylated genes are highlighted in blue, while hypermethylated genes are highlighted in red. IGV browser snapshots (bottom) of m 6 A-eCLIP depicting the coverage and change in the peak height between the NT and IGF2BP3-depleted cells for MAT2A 3′ UTR are shown. All data are n ≥ 3 replicates represented as mean ± standard deviation (SD), compared by two-sided unpaired t test; * p < 0.05, ** p < 0.01, and *** p < 0.001. In case of missing or outlier values, the replicate was not reported. All experiments were repeated at least twice for consistency. All the western blots were repeated at least three times to report the changes, if any. Refer also to .

Article Snippet: MAT2A , Proteintech , 55309-1-AP; RRID: AB_2881303.

Techniques: Western Blot, Methylation, Control, Dot Blot, Modification, Staining, Enzyme-linked Immunosorbent Assay, Isolation, Quantitative Proteomics, Standard Deviation

Journal: Cell reports

Article Title: IGF2BP3 redirects glycolytic flux to promote one-carbon metabolism and RNA methylation

doi: 10.1016/j.celrep.2025.116330

Figure Lengend Snippet: (A) Western blot analysis of Lin− cells from Igf2bp3 del/del mice. Briefly, cells were isolated from mice with a germline deletion of Igf2bp3, transformed with MLL-Af4, and then subjected to transduction with MSCV-based constructs carrying the wild-type murine Igf2bp3. Proteins that were analyzed are Igf2bp3, Mat2a, Mat2b, and actin. (B) Cell growth, measured by CellTiter-Glo, over 4 days in Igf2bp3 del/del Lin− MLL-Af4 cells with enforced IGF2BP3 expression as above. Viability has been normalized to control cells; mean ± standard deviation (SD) ( n = 5); one-way ANOVA followed by Bonferroni’s multiple comparisons test; **** p < 0.0001. (C) Representative Seahorse XF extracellular acidification rate (ECAR) kinetic trace in cells described above ( n = 4). (D) Aggregate lactate efflux rates from Seahorse XF analysis in cells described above; two-sided unpaired t test; * p < 0.05 ( n = 4). (E) Colony formation assays from Lin− MLL-Af4 cells as described above; two-sided unpaired t test; ** p < 0.01 ( n = 2). (F) ELISA measurement of m 6 A modification on RNA isolated from Igf2bp3 del/del Lin− MLL-Af4 cells with enforced IGF2BP3 expression as above; two-sided unpaired t test; * p < 0.05 ( n = 3). (G) Percentage engraftment of CD45.2 Lin− cells in bone marrow from Igf2bp3 del/del mice transduced with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. (H) Quantitation of bone marrow count in mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. (I) Spleen weights of mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. (J) Quantitation of spleen cell count in mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. (K) Quantitation of bone marrow CD11b+ cell count in mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. (L) Quantitation of bone marrow Lin− cell count along with representative fluorescence-activated cell sorting (FACS) plots in mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. (M) Quantitation of bone marrow CD11b+cKit+ cell count in mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. (N) Quantitation of bone marrow LSK (Lin− cKit+Sca1−) cell count in mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. (O) Quantitation of bone marrow CD11b+Sca1− (potential LIC ) cell count in mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. (P) Seahorse XF ECAR kinetic trace for bone marrow cells isolated from the empty vector (Ctrl) or IGF2BP3 re-expression group at 6 weeks ( n = 4, each group; for representation n = 2). (Q) Aggregate lactate efflux rates from Seahorse XF analysis in cells described above; reported as mean ± SD ( n = 4). (R) ELISA measurement of m 6 A RNA modifications in splenic tumors isolated from mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks; reported as mean ± SD; 8 mice/group. (S) H&E staining of spleen of mice transplanted with mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups at 6 weeks. Scale bar: 100 μm. (T) Overall survival of mice transplanted with MLL-Af4 re-expressing empty vector (Ctrl) or IGF2BP3 in the two groups (representative graph cumulative of two experiments, 8 mice/group; the experiment was terminated after 12 weeks; Kaplan-Meier method with log rank test was used to report the results). The animal experiments were repeated twice. All the western blots were repeated at least three times to report the changes, if any. Data in this figure are represented as mean ± SD with n = 8 mice per group. Statistical tests were performed using two-sided unpaired t test with significance levels as indicated; * p < 0.05, ** p < 0.01, and *** p < 0.001. Refer also to and .

Article Snippet: MAT2A , Proteintech , 55309-1-AP; RRID: AB_2881303.

Techniques: Western Blot, Isolation, Transformation Assay, Transduction, Construct, Expressing, Control, Standard Deviation, Enzyme-linked Immunosorbent Assay, Modification, Plasmid Preparation, Quantitation Assay, Cell Counting, Fluorescence, FACS, Staining