Journal: The FASEB Journal

Article Title: Leptin promotes methionine adenosyltransferase 2A expression in hepatic stellate cells by the downregulation of E2F‐4 via the β‐catenin pathway

doi: 10.1096/fj.201903021rr

Figure Lengend Snippet: FIGURE 1 Knockdown of MAT2A inhibits leptin-induced HSC activation and liver fibrosis. A, Western blot analyses of the protein levels of MAT2A, α1(I)collagen, and α-SMA (n = 3). HSCs in the 6-well plate were transfected with 1 μg of MAT2A siRNA or the control siRNA (the scrambled RNA) and underwent starvation before leptin treatment for 24 hours. The protein levels of MAT2A, α1(I)collagen, and α-SMA were examined by western blot analyses. The densities of the bands were expressed as fold change relative to the control. *P < .05. B, Fluorescence staining of α-SMA (a marker for activated HSCs), Sirius red staining of collagen in the liver, and western blot analysis of liver MAT2A. Two groups of ob/ob mice were injected with AAV.MAT2A siRNA (MAT2A siRNA, 5 × 1011 VG/mouse) or AAV.scrambled siRNA (Control siRNA) by tail vein after 2-week of treatment with TAA plus leptin and then received TAA plus leptin for additional 2-week. Fluorescence staining and Sirius red staining were performed for the respective examination of α-SMA level and collagen level in the liver and liver MAT2A protein levels were examined by western blot analysis. The activated HSCs (green) were counted in six randomly chosen fields at 200-fold magnification (Scale bar: 50 μm) and the fold changes were shown. The areas of red color (fibrosis area, Scale bar: 50 μm) were expressed as the fold change relative to the control. *P < .05

Article Snippet: For knockdown of the gene, HSCs in six-well plastic plates were transfected with 1 μg of small interferencing RNA (siRNA) targeting MAT2A (MAT IIα siRNA, Santa Cruz, CA, USA) or the control scrambled RNA (Control siRNA-A, Santa Cruz, CA, USA) by siRNA Transfection Reagent (Santa Cruz, CA, USA).

Techniques: Knockdown, Activation Assay, Western Blot, Transfection, Control, Fluorescence, Staining, Marker, Injection

Journal: The FASEB Journal

Article Title: Leptin promotes methionine adenosyltransferase 2A expression in hepatic stellate cells by the downregulation of E2F‐4 via the β‐catenin pathway

doi: 10.1096/fj.201903021rr

Figure Lengend Snippet: FIGURE 2 Leptin upregulates MAT2A expression in HSCs. A, B, Western blot and real-time PCR analyses of MAT2A expression (n = 3). Serum-starved HSCs were stimulated with different doses of leptin for 24 hours and the MAT2A levels of protein and mRNA were evaluated by western blot (A) and real-time PCR (B), respectively. The data are representative of three independent experiments. The densities of the bands from western blot were expressed as fold change relative to the control. *P < .05. C, Transfection assay for analysis of MAT2A promoter activity (n = 3). HSCs transfected with 1.6 μg of pGL4MAT2A(−2954)Luc were starved and incubated with or without leptin for 24 hours. Luciferase assays were performed. *P < .05. D, Double fluorescence staining for examining MAT2A in HSCs in the ob/ob mouse model. Two groups of ob/ ob mice were treated with or without leptin (1 μg/g body weight, once per day) throughout 4-week period of treatment with TAA (200 μg/g body weight, three times a week) by i.p.. Double fluorescence staining was performed on the sections of liver tissue. The nuclei were counterstained with Hoechst 33342. The activated HSCs (green) and the MAT2A-positive activated HSCs (yellow) were counted in six randomly chosen fields at 200- fold magnification and the fold changes were shown. Scale bar: 50 μm. *P < .05

Article Snippet: For knockdown of the gene, HSCs in six-well plastic plates were transfected with 1 μg of small interferencing RNA (siRNA) targeting MAT2A (MAT IIα siRNA, Santa Cruz, CA, USA) or the control scrambled RNA (Control siRNA-A, Santa Cruz, CA, USA) by siRNA Transfection Reagent (Santa Cruz, CA, USA).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Control, Transfection, Activity Assay, Incubation, Luciferase, Fluorescence, Staining

Journal: The FASEB Journal

Article Title: Leptin promotes methionine adenosyltransferase 2A expression in hepatic stellate cells by the downregulation of E2F‐4 via the β‐catenin pathway

doi: 10.1096/fj.201903021rr

Figure Lengend Snippet: FIGURE 3 β-Catenin mediates leptin upregulation of MAT2A expression in HSCs. A, B, Western blot and real-time PCR analyses of MAT2A expressions (n = 3). HSCs were undergone starvation and then treated with or without 5 μM of XAV939 before leptin stimulation for another 24 hours. MAT2A expressions were examined by western blot and real-time PCR. The densities of the bands from western blot were expressed as the fold change. *P < .05. C, Transfection assay for analysis of MAT2A promoter activity (n = 3). HSCs transfected with 1.6 μg of pGL4MAT2A(−2954)Luc were starved and incubated with or without 5 μM of XAV939 before leptin treatment for another 24 hours. Luciferase assays were performed. *P < .05. D, E, F, Double fluorescence staining for examining MAT2A and β-catenin in activated HSCs in the ob/ob mouse model and Sirius red staining for evaluating collagen level. Ob/ob mice were treated with XAV939 (5 μg/g body weight, once per day) or the vehicle throughout 4-week administration of TAA (200 μg/g body weight, three times a week) plus leptin (1 μg/g body weight, once per day) or TAA plus vehicle. Double fluorescence staining was conducted on the sections of liver tissue for detecting the expression of MAT2A and β-catenin in HSCs. The nuclei were counterstained with Hoechst 33342. The numbers of the activated HSCs (green in D; red in E), MAT2A-positive activated HSCs (yellow), and β-catenin-positive activated HSCs (yellow) were counted in six randomly chosen fields at 200-fold magnification and the fold changes were shown (Scale bar: 50 μm). Sirius red was also performed by the liver sections for evaluating collagen level (F) and the red areas were expressed as the fold change relative to the control (Scale bar: 100 μm). *P < .05

Article Snippet: For knockdown of the gene, HSCs in six-well plastic plates were transfected with 1 μg of small interferencing RNA (siRNA) targeting MAT2A (MAT IIα siRNA, Santa Cruz, CA, USA) or the control scrambled RNA (Control siRNA-A, Santa Cruz, CA, USA) by siRNA Transfection Reagent (Santa Cruz, CA, USA).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Transfection, Activity Assay, Incubation, Luciferase, Fluorescence, Staining, Control

Journal: The FASEB Journal

Article Title: Leptin promotes methionine adenosyltransferase 2A expression in hepatic stellate cells by the downregulation of E2F‐4 via the β‐catenin pathway

doi: 10.1096/fj.201903021rr

Figure Lengend Snippet: FIGURE 4 Leptin affects MAT2A promoter activity by elements between position −2847 bp and −2752 bp in MAT2A promoter. Two groups of HSCs were transfected, respectively, with the promoter report plasmids harboring different lengths of MAT2A promoter (pMAT2A(−2954)Luc, pMAT2A(−2242)Luc, pMAT2A(−2073)Luc, pMAT2A(−2847)Luc, pMAT2A(−2752)Luc, pMAT2A(−2655)Luc, pMAT2A(−2470)Luc) (A,B). The third group of HSCs was transfected with pMAT2A(−2847)Luc or pMAT2A(E2F4 mut)Luc (The possible E2F-4 binding site was mutated by pMAT2A (−2847)Luc)(C). All the transfected HSCs were starved before treatment with leptin for 24 hours. The fourth group of HSCs was cotransfected with pMAT2A(−2847)Luc or pMAT2A(E2F4 mut)Luc plus pcDNAE2F4 or the empty plasmids (pcDNA3.1) and then incubated for 24 hours (D). Luciferase assays were performed. The data were from three independent experiments. *P < .05

Article Snippet: For knockdown of the gene, HSCs in six-well plastic plates were transfected with 1 μg of small interferencing RNA (siRNA) targeting MAT2A (MAT IIα siRNA, Santa Cruz, CA, USA) or the control scrambled RNA (Control siRNA-A, Santa Cruz, CA, USA) by siRNA Transfection Reagent (Santa Cruz, CA, USA).

Techniques: Activity Assay, Transfection, Binding Assay, Incubation, Luciferase

Journal: The FASEB Journal

Article Title: Leptin promotes methionine adenosyltransferase 2A expression in hepatic stellate cells by the downregulation of E2F‐4 via the β‐catenin pathway

doi: 10.1096/fj.201903021rr

Figure Lengend Snippet: FIGURE 5 E2F-4 binds to MAT2A promoter around a site at −2779 bp, which is attenuated by leptin stimulation. A, EMSA of E2F-4 binding to a possible site around a position −2779 bp in MAT2A promoter. Nuclear extracts (NE) were prepared from HSCs stimulated with leptin (L) or vehicle (V). About 6 μg of nuclear proteins was incubated with biotinylated DNA fragment containing the potential E2F-4 binding site around −2779 bp. For competition assay, 6 μg of nuclear proteins was preincubated with 200-fold molar excess of the unlabeled probe before the addition of the labeled probe. For supershift assay, 6 μg of nuclear proteins was preincubated with 2 μg of anti-E2F-4 antibody before the addition of the labeled probe. B, ChIP analysis of the interaction between E2F-4 and the possible E2F-4 binding site in MAT2A promoter. HSCs were incubated with or without leptin (Lep) for 24 hours and ChIP analyses were conducted by anti-E2F-4 antibody. The fragment (109 bp) containing the possible E2F-4 binding site in MAT2A promoter was detected by PCR. The PCR products were subjected to electrophoresis in 2% agarose gel. The representative results were shown from three independent experiments

Article Snippet: For knockdown of the gene, HSCs in six-well plastic plates were transfected with 1 μg of small interferencing RNA (siRNA) targeting MAT2A (MAT IIα siRNA, Santa Cruz, CA, USA) or the control scrambled RNA (Control siRNA-A, Santa Cruz, CA, USA) by siRNA Transfection Reagent (Santa Cruz, CA, USA).

Techniques: Binding Assay, Incubation, Competitive Binding Assay, Labeling, Electrophoresis, Agarose Gel Electrophoresis

Journal: EBioMedicine

Article Title: TGF-β1/p65/MAT2A pathway regulates liver fibrogenesis via intracellular SAM

doi: 10.1016/j.ebiom.2019.03.058

Figure Lengend Snippet: Upregulation of MAT2A reduced SAM concentration in the fibrotic liver tissues. (a) Representative Western blot results of MAT2A, TGF-β1, COL1A1 and α-SMA in mice liver tissues from Control (Con), CCl 4 treatment (CCl 4 ) or CCl 4 combined with NPLC0393 treatment (CCl 4 + NPLC0393) and NPLC0393 alone treatment (NPLC0393) groups. The densitometric changes in fold change over Control group were shown in right panel ( n = 6 per group). (b) Immunofluorescence staining of MAT2A (green) and the HSC marker α-SMA (red) in mice liver tissues. Co-localization of MAT2A with α-SMA was shown in yellow (n = 6 per group). Scale bar, 100 μm. (c) qRT-PCR result of mat2a, a-sma and col1a1 mRNA expressions in mice liver tissues from each group. Gene expression was normalized using gapdh as the calibrator gene and fold changes over Control group were shown. Data are presented as means ± S.D. * P < .05, ** P < .01 and *** P < .001 compared with Control group; # P < .05, ## P < .01 ### P < .001 compared with CCl 4 group ( n = 6 per group). (d) The SAM concentration in mice liver tissues from control, CCl 4 model and NPLC0393 treatment groups were measured ( n = 5 per group). Data are presented as means ± S.D. * P < .05, ** P < .01 and *** P < .001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The siRNA targeting human MAT2A ( MAT2A siRNA-1: 5′- GUGAGAGAGAGCUAUUAGA-3′, MAT2A siRNA-2: 5′-ACACAUUGGAUAUGAUGAU-3′, MAT2A siRNA-3: 5′- AGCAGUUGUGCCUGCGAAA-3′), human PPM1A/PP2Cα (5′- GAGUUAUGUCAGAGAAGAA-3′) were synthesized by Genepharma Co. (Shanghai, China).

Techniques: Concentration Assay, Western Blot, Control, Immunofluorescence, Staining, Marker, Quantitative RT-PCR, Gene Expression

Journal: EBioMedicine

Article Title: TGF-β1/p65/MAT2A pathway regulates liver fibrogenesis via intracellular SAM

doi: 10.1016/j.ebiom.2019.03.058

Figure Lengend Snippet: Upregulation of MAT2A led to the decrease of SAM concentration in the TGF-β1-induced activated HSCs. (a) The normal human hepatocyte cell line L02 and human HSC cell line LX-2 were treated with or without 10 μM NPLC0393 for 24 h and then stimulated with 5 ng/mL TGF-β1 for another 24 h. The levels of MAT2A, COL1A1 and α-SMA proteins were measured by Western blot. Data are representative of three independent experiments. (b) LX-2 cells were treated as in (a) and the expression level of MAT2A, COL1A1 and α-SMA mRNA were examined by qRT-PCR. * P < .05, ** P < .01 and *** P < .001 compared with Control group; # P < .05, ## P < .01 ### P < .001 compared with TGF-β1 treatment group ( n = 3 per group with duplicates). (c) The SAM concentration in LX-2 cells from Control, TGF-β1 and NPLC0393 treatment groups were measured by LC-MS (n = 6 per group with duplicates). * P < .05, ** P < .01. (d) LX-2 cells were pre-treated with 2 mM SAM or 10 μM NPLC0393 for 24 h and then stimulated with 5 ng/mL TGF-β1 in the complete DMEM or L-methionine-free DMEM for another 24 h. The expression levels of MAT2A, MAT2B, COL1A1 and α-SMA proteins were analyzed by Western blot. Met (+) means normal DMEM culture medium and Met (−) means L-methionine-free DMEM culture medium. Data are representative of three independent experiments.

Article Snippet: The siRNA targeting human MAT2A ( MAT2A siRNA-1: 5′- GUGAGAGAGAGCUAUUAGA-3′, MAT2A siRNA-2: 5′-ACACAUUGGAUAUGAUGAU-3′, MAT2A siRNA-3: 5′- AGCAGUUGUGCCUGCGAAA-3′), human PPM1A/PP2Cα (5′- GAGUUAUGUCAGAGAAGAA-3′) were synthesized by Genepharma Co. (Shanghai, China).

Techniques: Concentration Assay, Western Blot, Expressing, Quantitative RT-PCR, Control, Liquid Chromatography with Mass Spectroscopy

Journal: EBioMedicine

Article Title: TGF-β1/p65/MAT2A pathway regulates liver fibrogenesis via intracellular SAM

doi: 10.1016/j.ebiom.2019.03.058

Figure Lengend Snippet: Profibrotic effect of MAT2A on liver fibrogenesis. (a) Mice were injected via tail vein with AAV- MAT2A or AAV-empty vector (AAV-EV) for 3 weeks and subjected to CCl 4 administration with or without NPLC0393 for another 4 weeks. Representative image of MAT2A fluorescence, Sirius Red and α-SMA staining of the liver tissues were shown (Scale bars, 100 μm, n = 6 per group). (b) Semi-quantitative scoring of α-SMA staining for each group was shown in a bar graph. IOD: integrated optical density. (c) Representative Western blot result of Flag-MAT2A, α-SMA and COL1A1 proteins in mice livers from different groups. (d) The SAM concentration in mice liver from different groups were measured by LC-MS. * P < .05, ** P < .01 (n = 6 per group). (e) LX-2 cells transfected with the Negative Control siRNA (NC siRNA) or MAT2A siRNA were treated with 5 ng/mL TGF-β1 for 24 h. The expression levels of MAT2A, MAT2B, α-SMA and COL1A1 proteins were analyzed by Western blot. Data are representative of three independent experiments. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The siRNA targeting human MAT2A ( MAT2A siRNA-1: 5′- GUGAGAGAGAGCUAUUAGA-3′, MAT2A siRNA-2: 5′-ACACAUUGGAUAUGAUGAU-3′, MAT2A siRNA-3: 5′- AGCAGUUGUGCCUGCGAAA-3′), human PPM1A/PP2Cα (5′- GAGUUAUGUCAGAGAAGAA-3′) were synthesized by Genepharma Co. (Shanghai, China).

Techniques: Injection, Plasmid Preparation, Fluorescence, Staining, Western Blot, Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Transfection, Negative Control, Expressing

Journal: EBioMedicine

Article Title: TGF-β1/p65/MAT2A pathway regulates liver fibrogenesis via intracellular SAM

doi: 10.1016/j.ebiom.2019.03.058

Figure Lengend Snippet: Phosphorylation of p65 mediated TGF-β1-induced MAT2A expression. (a) Representative Western blot result of the total p65 and phospho-p65 (p-p65) in the control, CCl 4 -treated or NPLC0393-treated mice (n = 6 per group). (b) LX-2 cells were pre-treated with 10 μM NPLC0393 or 10 μM SB431542 for 24 h and then stimulated with 5 ng/mL TGF-β1 for another 2 h. The expression levels of the total p65 and p-p65 were analyzed by Western blot. (c) LX-2 cotransfected with the 1 μg/mL NF-κB reporter gene and 1 μg/mL pRL-TK plasmid were treated with vehicle, NPLC0393 or 10 μM SB431542 for 12 h and then stimulated with 5 ng/mL TGF-β1 for another 24 h. The Luciferase activity of NF-κB was normalized against that of the cotransfected pRL-TK. The result was shown as fold changes over untreated transfectants obtained from three independent tests. (d) LX-2 cells cotransfected with the 1 μg/mL MAT2A promoter construct and 1 μg/mL pRL-TK plasmid were treated with NPLC0393 or 50 ng/mL SN50 for 12 h and then stimulated with 5 ng/mL TGF-β1 for another 24 h. Relative luciferase activities were analyzed as above. (e) LX-2 cells infected with Ad-CMV-p65 (Ad-p65) or Ad-empty vector (Ad-EV) were treated with or without NPLC0393 for 12 h and then stimulated with 5 ng/mL TGF-β1 for another 24 h and the expression levels of p-p65, MAT2A, COL1A1 and α-SMA proteins were analyzed by Western blot. (f) LX-2 cells transfected with NC siRNA or PP2Cα siRNA were treated as (e) and the expression levels of p-p65, MAT2A and α-SMA proteins were analyzed by Western blot. * P < .05, ** P < .01 and *** P < .001. Data are representative of three independent experiments.

Article Snippet: The siRNA targeting human MAT2A ( MAT2A siRNA-1: 5′- GUGAGAGAGAGCUAUUAGA-3′, MAT2A siRNA-2: 5′-ACACAUUGGAUAUGAUGAU-3′, MAT2A siRNA-3: 5′- AGCAGUUGUGCCUGCGAAA-3′), human PPM1A/PP2Cα (5′- GAGUUAUGUCAGAGAAGAA-3′) were synthesized by Genepharma Co. (Shanghai, China).

Techniques: Phospho-proteomics, Expressing, Western Blot, Control, Plasmid Preparation, Luciferase, Activity Assay, Construct, Infection, Transfection

Journal: EBioMedicine

Article Title: TGF-β1/p65/MAT2A pathway regulates liver fibrogenesis via intracellular SAM

doi: 10.1016/j.ebiom.2019.03.058

Figure Lengend Snippet: Schematic representation of TGF-β1/p65/MAT2A signaling pathway and inhibitory effect of NPLC0393 on this pathway. In HSCs, TGF-β1 induces MAT2A expression via p65 phosphorylation, which results in decrease of SAM concentration and consequential increase of α-SMA and COL1A1 expressions. PP2Cα small molecular activator NPLC0393 downregulates MAT2A by inhibiting phosphorylation of p65, thereby maintaining intracellular SAM concentration.

Article Snippet: The siRNA targeting human MAT2A ( MAT2A siRNA-1: 5′- GUGAGAGAGAGCUAUUAGA-3′, MAT2A siRNA-2: 5′-ACACAUUGGAUAUGAUGAU-3′, MAT2A siRNA-3: 5′- AGCAGUUGUGCCUGCGAAA-3′), human PPM1A/PP2Cα (5′- GAGUUAUGUCAGAGAAGAA-3′) were synthesized by Genepharma Co. (Shanghai, China).

Techniques: Expressing, Phospho-proteomics, Concentration Assay

Journal: Cell & Bioscience

Article Title: mTORC1-c-Myc pathway rewires methionine metabolism for HCC progression through suppressing SIRT4 mediated ADP ribosylation of MAT2A

doi: 10.1186/s13578-022-00919-y

Figure Lengend Snippet: SIRT4 regulates methionine metabolism through mediating MAT2A ADP ribosylation. A Volcano plot showing differential gene expression in mice’s livers upon methionine restriction. Red circles indicates genes increased in the Ctrl set while blue ones means genes decreased in the MR set. B Western blot analysis of MAT2A mono-ADP-ribosylation (MARylation) in HEK293 and SNU449 cells in which SIRT4 was silenced (shRNA). C The MARylation of MAT2A was measured by western blot in HEK293 or SNU449 cells stably expressing HA tagged SIRT4. D Western blot analysis of MAT2A MARylation in the FLAG immunoprecipitated samples from HEK293 or SNU449 cells treated with nicotinamide (NAM) for indicated times. E Western blot analysis of MAT2A MARylation in the FLAG immunoprecipitated samples from HEK293 or SNU449 cells treated with Sirtinol for indicated times. F Relative activity of MAT2A was shown in the SNU449 cells in which either c-Myc or SIRT4 was knockdown or overexpressed or c-Myc and SIRT4 overexpressed together. G Abundances of intracellular primary methionine cycle metabolites were compared between the control cells and the SNU449 cells where either c-Myc or SIRT4 was knockdown or stably overexpressed or c-Myc and SIRT4 overexpressed together. H MARylation level of mutant FLAG-MAT2A-E111A ectopically expressed in MAT2A-knockout HEK293 or SNU449 cells was measured by using anti-ADP ribosylation antibody when SIRT4 was silenced. I MARylation level of mutant FLAG-MAT2A-E111A ectopically expressed in MAT2A-knockout HEK293 or SNU449 cells was measured by using anti-ADP ribosylation antibody when SIRT4 was overexpressed. J MAT2A dimer are shown in blue or lightblue (PDB ID: 4NDN). The active site was shown in red circle. K E111 in MAT2A is conserved. Protein sequence alignment surrounding E111 colored in light blue from indicated species. Data are means ± SEM. Group differences were analyzed by two-tailed Student’s multiple comparison test ( F , G ) (**p < 0.01)

Article Snippet: c-Myc shRNA (sc-29226-SH), TRIM32 siRNA (sc-61714), TRIM32 shRNA (sc-61714-SH), SIRT4 shRNA (sc-63024-SH), MAT2A shRNA (sc-106203-SH) were purchased from Santa Cruz Biotechnology.

Techniques: Gene Expression, Western Blot, shRNA, Stable Transfection, Expressing, Immunoprecipitation, Activity Assay, Knockdown, Control, Mutagenesis, Knock-Out, Sequencing, Two Tailed Test, Comparison

Journal: Cell & Bioscience

Article Title: mTORC1-c-Myc pathway rewires methionine metabolism for HCC progression through suppressing SIRT4 mediated ADP ribosylation of MAT2A

doi: 10.1186/s13578-022-00919-y

Figure Lengend Snippet: Myc promoted SIRT4 degradation to reduce ADP ribosylation of MAT2A in the HCC. A Protein levels of c-Myc, MAT2A and MARylation level of MAT2A were determined by western blot from 85 pairs of tumour tissues. In the inserted boxplots, the circles indicate the median; the square indicate the 5th–95th percentiles. The statistical analysis was performed by the two tailed Student’s t-test (**p < 0.01). B Correlation between Myc and SIRT4 in HCC patients. 85 tissue pairs were immunohistochemically stained with indicated antibodies. The pair-wise Pearson correlation coefficient and the corresponding p-value between two genes were calculated. C The patients with high SIRT4 expression (n = 43) have longer survival compared to low SIRT4 expression (n = 42). Significance was determined using Kaplan–Meier analysis. D Schematic showing that high methionine content is sensed by mTORC1 that triggers c-Myc expression, which promotes TRIM32-mediated SIRT4 degradation. SIRT4 regulates MAT2A activity through MARylation, thereby inhibits MAT2A mediated tumour growth. SIRT4 loss resulted in the low MARylation level of MAT2A, which promotes methionine metabolism to generate SAM. In a feedback, high SAM level triggers transcription of metabolic enzymes to maintain tumour progression. Upward pointing arrows indicate an increase, and downwards pointing arrows indicate a decrease

Article Snippet: c-Myc shRNA (sc-29226-SH), TRIM32 siRNA (sc-61714), TRIM32 shRNA (sc-61714-SH), SIRT4 shRNA (sc-63024-SH), MAT2A shRNA (sc-106203-SH) were purchased from Santa Cruz Biotechnology.

Techniques: Western Blot, Two Tailed Test, Staining, Expressing, Activity Assay