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: The catalytic efficiency of METTL16 affects cellular processes by governing the intracellular S-adenosylmethionine setpoint

doi: 10.1016/j.celrep.2025.115966

Figure Lengend Snippet: (A) Two-tiered model of METTL16-mediated regulation of MAT2A RNA. Red hp denotes methylation. (B) Representative western blot of protein from METTL16-AID cell line after auxin treatment. In all figures herein positions of molecular weight markers are given in kDa to the right of the panel. Quantification of METTL16 protein normalized to PABPN1 for loading control, with time 0 set to 1 ( n = 3). (C) Cell growth was measured using CellTiter Glo in parental HCT116 and METTL16-AID cells after auxin treatment. Day 6 −auxin is set to 1 ( n = 3). (D) Northern blot of total RNA from METTL16-AID cells after ±auxin for 2 h followed by Met depletion. GAPDH is a loading control. MAT2A quantified as percent detained intron. One-way ANOVA comparing all samples to 0 h +auxin was performed. *** p = 0.0001, ns p = 0.0761, **** p < 0.0001 ( n = 3). (E) Northern blot of total RNA from METTL16-AID cells after auxin treatment or overnight (O/N). MAT2A mRNA levels were normalized to GAPDH and time 0 was set to 1. Unpaired two-tailed t tests with Welch’s correction comparing each time point to 0 h were performed. p = **0.0098 (2 h), **0.0012 (6 h), ***0.0003, and *0.0418 ( n = 3). (F) SAM levels measured by HPLC-tandem mass spectrometry from EV cells after auxin treatment. SAM levels were normalized to total ion count (TIC) and the average of −auxin samples was set to 1. Unpaired two-tailed t tests comparing − with + at each time point were performed ( n ≥ 3). All error bars represent standard deviation from the mean.

Article Snippet: MAT2A , Novus , Cat#NB110-94158; RRID: AB_1237164.

Techniques: Methylation, Western Blot, Molecular Weight, Control, Northern Blot, Two Tailed Test, Mass Spectrometry, Standard Deviation

Journal: Cell reports

Article Title: The catalytic efficiency of METTL16 affects cellular processes by governing the intracellular S-adenosylmethionine setpoint

doi: 10.1016/j.celrep.2025.115966

Figure Lengend Snippet: (A) CellTiter Glo (CTG) assays in complemented METTL16-AID cells after auxin treatment. Day 0 is set to 1 ( n = 2). (B) Northern blot of total RNA after 24 h of auxin and/or MAT2Ai treatment in EV, WT, or K163A cells. GAPDH is a loading control. One-way ANOVAs comparing the +auxin samples with WT+ for both − and + MAT2Ai were performed. *** p = 0.0009 (EV+ to WT+, − MAT2Ai), ** p = 0.0020, **** p < 0.0001, and *** p = 0.0007 (WT+ to K163A+, +MAT2Ai) ( n = 3). (C) SAM levels measured by HPLC-tandem mass spectrometry ±auxin for 48 h in EV, WT, or K163A cells. SAM levels were normalized to total ion count (TIC) for loading control, with EV −auxin set to 1. One-way ANOVA comparing the +auxin samples with WT+ was performed. p = 0.0004 and p = 0.0007 ( n = 3). The data for EV are from the 48 h data shown in . (D) CTG assays in WT or K163A complemented cells after auxin treatment. Cells were conditioned in ±auxin medium for 5–7 days prior to seeding. Day 0 is set to 1 ( n = 3). Unpaired two-tailed t tests were performed between K163A and WT +auxin. For day 4, p = 0.0291. For day 5, p = 0.0039. (E) CTG assays of WT or K163A cells after auxin and MAT2Ai treatment. Five days at 0 nM MAT2Ai is set to 1 for each cell line ( n = 3). (F) Western blot of protein from WT or K163A cells after 5 days of auxin treatment with MAT2Ai. MAT2A protein was normalized to actin and WT, 1,000 nM MAT2Ai was set to 1 ( n = 2). Protein was harvested from the same cells used on day 5 of the CTG assays in (E). (G) SAM levels measured by HPLC-tandem mass spectrometry after auxin and/or MAT2Ai treatment for 5 days in WT or K163A cells. SAM levels were normalized to TIC for loading. The −auxin/−MAT2Ai sample was set to 1 for each cell line. Unpaired two-tailed t test comparing WT with K163A for +auxin/−MAT2Ai was performed ( n = 3). (H) Same data as (G) but omitting the −MAT2Ai samples for better visualization of +MAT2Ai data. All error bars represent standard deviation from the mean.

Article Snippet: MAT2A , Novus , Cat#NB110-94158; RRID: AB_1237164.

Techniques: Northern Blot, Control, Mass Spectrometry, Two Tailed Test, Western Blot, Standard Deviation

Journal: Cell reports

Article Title: The catalytic efficiency of METTL16 affects cellular processes by governing the intracellular S-adenosylmethionine setpoint

doi: 10.1016/j.celrep.2025.115966

Figure Lengend Snippet: (A) Western blots of total protein from EV, K163A, WT, GFP, or MAT2A (M2A) cells after 48 h of auxin and/or MAT2Ai treatment. Protein was run in parallel for H3K4me3, H3K36me3, H3K9me3, H3K27me3, and MAT2A. Actin, H3, and PABPN1 are loading controls. (B) MAT2A protein levels were normalized to actin, with −auxin, +MAT2Ai set to 1. Unpaired two-tailed t tests were performed for the indicated comparisons ( n = 3). (C) H3K4me3 protein levels were normalized to actin, with −auxin, −MAT2Ai set to 1, except for GFP and MAT2A in which −auxin, +MAT2Ai was set to 1. Unpaired two-tailed t tests were performed for the indicated comparisons, with p values noted in the figure ( n = 3). (D) Quantification of H3K36me3 levels as in (C) ( n = 3). (E) Quantification of H3K9me3 levels as in (C) but normalized to H3 ( n = 3). (F) Quantification of H3K27me3 levels as in (C) but normalized to PABPN1 ( n = 3). All error bars represent standard deviation from the mean.

Article Snippet: MAT2A , Novus , Cat#NB110-94158; RRID: AB_1237164.

Techniques: Western Blot, Two Tailed Test, Standard Deviation

Journal: Cell reports

Article Title: The catalytic efficiency of METTL16 affects cellular processes by governing the intracellular S-adenosylmethionine setpoint

doi: 10.1016/j.celrep.2025.115966

Figure Lengend Snippet: (A) SUnSET assay (left) and quantification (right) of puromycin signal from METTL16-AID cells after auxin treatment followed by 15 min of puromycin treatment. Cycloheximide (CHX) was used as a negative control. Puromycin-labeled proteins were normalized to PABPN1, with −auxin set to 1. Unpaired two-tailed t tests were performed for the indicated comparisons, using Welch’s correction when a sample was set to 1 ( n = 3). (B) SUnSET assay as in (A) using GFP and MAT2A complemented METTL16-AID cells after 48 h of auxin and MAT2Ai treatment followed by 15 min of puromycin. Puromycin-labeled protein levels were normalized to PABPN1, with −auxin, +MAT2Ai for each cell line set to 1. Unpaired two-tailed t tests were performed for the indicated comparisons, using Welch’s correction when a sample was set to 1, with p values noted in the figure ( n = 3). Both images are from the same blot and brightness was adjusted prior to cropping. All error bars represent standard deviation from the mean.

Article Snippet: MAT2A , Novus , Cat#NB110-94158; RRID: AB_1237164.

Techniques: Negative Control, Labeling, Two Tailed Test, Standard Deviation

Journal: Cell reports

Article Title: The catalytic efficiency of METTL16 affects cellular processes by governing the intracellular S-adenosylmethionine setpoint

doi: 10.1016/j.celrep.2025.115966

Figure Lengend Snippet: (A) Number of hypomethylated or hypermethylated sites identified in the five comparison groups. (B) Percent of m 6 A sites within each consensus motif for all or hypomethylated sites. Numbers above the bars represent number of sites. (C) Metagene plot of all m 6 A sites identified in the EV+/WT+ samples. (D) Metagene plot of the hypomethylated sites in the EV+/WT+ samples. (E) The fold change (Log2FC) of the 61 U6 snRNA pseudogenes (RNU6) identified in any of the five comparison groups. One-way ANOVA with Welch’s correction was performed to determine p values. (F) The fold change of each hypomethylated U6 snRNA pseudogene and all other hypomethylated transcripts (Other) within each comparison group. Unpaired t tests with Welch’s correction were performed between each pair. (G) The m 6 A abundance in EV-auxin conditions is shown for all hypomethylated RNAs in the EV+/EV− comparison group. U6 snRNA pseudogenes (RNU6) were compared with all other transcripts (Other) ±auxin. MAT2A hp1 and hp3 sites are blue. Unpaired t tests with Welch’s correction were performed to determine significance of the differences between the −auxin samples. (H) Alignment of the consensus TACAGAGAA sequence variants in 24 of 61 U6 snRNA pseudogenes. Blue background with white letters conforms to the consensus. (I) The m 6 A abundance for each of the three −auxin samples for the 61 U6 snRNA pseudogenes according to the consensus variant. Variant bases are underlined. An unpaired t test with Welch’s correction was performed to determine the significance of the difference between the consensus and most common variant TATAGAGAA.

Article Snippet: MAT2A , Novus , Cat#NB110-94158; RRID: AB_1237164.

Techniques: Comparison, Sequencing, Variant Assay

Journal: Cell reports

Article Title: The catalytic efficiency of METTL16 affects cellular processes by governing the intracellular S-adenosylmethionine setpoint

doi: 10.1016/j.celrep.2025.115966

Figure Lengend Snippet: (A) CellTiter Glo assays of WT or K163A cells with MTAP deletion or NT controls. Each point was quantified relative to the matched sgNT ( n = 4 except 12 days n = 3). These values for K163A lines were compared with WT by two-tailed unpaired t test. For all three sgMTAP samples, ** p < 0.01 at 9 and 12 days. (B) Western blot from WT or K163A cells with MTAP-deleted or NT controls after 48 h of auxin. Protein was run in duplicate for MAT2A and SDMA, with actin loading controls shown for each blot. (C) Quantification of western blots as in (B). MAT2A was normalized to actin, with sgNT in WT cells set to 1 ( n = 3). Unpaired two-tailed t tests compared K163A plus auxin samples to their corresponding WT controls. p values for sgNT, sgMTAP1, sgMTAP2, and sgMTAP3 were ***0.00045, *0.017, **0.0057, and **0.0068, respectively. (D) Quantification of western blots as in (B). SDMA protein levels from the 50–75 kDa cluster were normalized to actin, with sgNT for each cell line set to 1 ( n = 3). All error bars represent standard deviation from the mean.

Article Snippet: MAT2A , Novus , Cat#NB110-94158; RRID: AB_1237164.

Techniques: Two Tailed Test, Western Blot, Standard Deviation