norepinephrine bitartrate monohydrate na injection experiment  (MedChemExpress)

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: To interfere with intracellular SAM levels, cells were cultured in complete medium containing freshly reconstituted 200 μM SAM (A7007, Sigma-Aldrich, St. Louis, MO, USA) for 48 h. To investigate the effect of norepinephrine, cells were treated with 10 μM norepinephrine (HY-137158, MedChemExpress, New Jersey, USA) for 24 h. The compound was dissolved in DMSO (D8370, Solarbio, Beijing, China) and then diluted in culture medium to the final concentration.

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: To interfere with intracellular SAM levels, cells were cultured in complete medium containing freshly reconstituted 200 μM SAM (A7007, Sigma-Aldrich, St. Louis, MO, USA) for 48 h. To investigate the effect of norepinephrine, cells were treated with 10 μM norepinephrine (HY-137158, MedChemExpress, New Jersey, USA) for 24 h. The compound was dissolved in DMSO (D8370, Solarbio, Beijing, China) and then diluted in culture medium to the final concentration.

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

Journal: Advanced Science

Article Title: CARD9 Conveys Pancreatic Islet Sympathetic Nervous β2 Signals to Reshape Macrophage Creatine Metabolism in Type 1 Diabetes

doi: 10.1002/advs.202507543

Figure Lengend Snippet: CARD9 is a regulator for macrophage conveying of β2‐AR signaling. A) Flowchart illustrating the synthesis of neurotransmitters by sympathetic neurons. B) NE levels in the serum and pancreatic tissues of NOD mice at different ages. C) Scatter plot showing the correlation between NE levels and CD11c + macrophage populations in the serum of NOD mice. D) Relative transcription levels of NE receptors in pancreatic islet macrophages of NOD mice at different ages. E) Western blot analysis of ADRB2 (β2‐adrenergic receptor) expression in pancreatic islet macrophages of NOD mice at different ages. F) Functional enrichment analysis of differentially expressed genes in BMDMs treated with β2‐AR agonist formoterol (FMT) or β2‐AR inhibitor butoxamine (BTX) compared to controls. G) Gene module clustering dendrogram from weighted gene co‐expression network analysis (WGCNA), illustrating modules formed based on gene co‐expression patterns. H) Module‐trait correlation heatmap from WGCNA, showing the associations between gene modules and treatment. I) Module eigengene‐trait correlation plots from WGCNA, illustrating the relationships between module eigengenes and treatments (FMT and BTX). J) Venn diagrams displaying macrophage activation‐related genes that are inversely regulated by FMT and BTX treatments. K) Venn diagrams displaying genes regulated by macrophages, those co‐regulated by β2‐AR, genes within the “brown” module, and differential gene sets of macrophage subsets in NOD mice. L) qPCR analysis of CARD9 expression levels in BMDMs treated with BTX and FMT. M) Western blot analysis of CARD9 protein expression in BMDMs and RAW264.7 following BTX and FMT treatment. N) Functional analysis of macrophages stably transfected with CARD9 overexpression plasmids. The levels of CD11c and CD206 were measured under the F4/80 + gate after exogenous administration of FMT and BTX. O) Functional analysis of macrophages with stable CARD9 knockdown. The levels of CD11c and CD206 were measured under the F4/80 + gate following treatment with FMT and BTX. P) Schematic summary of the overall experimental concept and key findings. The data were compiled from at least three independent experiments and are presented as the means ± SEMs. * p < 0.05, ** p < 0.01, *** p < 0.001 by Student's unpaired two‐tailed t ‐test or one‐way analysis of variance (ANOVA) with Tukey's post hoc comparison.

Article Snippet: For β2‐AR studies, cells were pre‐treated with FMT (100 n m ) or BTX (10 μ m ) for 2 h and then treated with NE (100 n m ; HY‐13715, MedChemExpress, USA) for 12 h. [ , , ] The mouse CARD9 expression plasmids and sh Card9 were obtained from Gene Create (Wuhan, China).

Techniques: Western Blot, Expressing, Functional Assay, Activation Assay, Stable Transfection, Transfection, Over Expression, Knockdown, Two Tailed Test, Comparison

Journal: Advanced Science

Article Title: CARD9 Conveys Pancreatic Islet Sympathetic Nervous β2 Signals to Reshape Macrophage Creatine Metabolism in Type 1 Diabetes

doi: 10.1002/advs.202507543

Figure Lengend Snippet: β2‐AR signaling regulates macrophage activation via the PKA/CREB1/CARD9 axis. A) Analysis of the overlap between β2‐AR‐regulated transcription factors and CARD9‐predicted transcription factors. B) Scatter plot showing the correlation between CARD9 and CREB1 expression levels in pancreatic tissues. C) Expression levels of Creb1 in BMDMs treated with BTX and FMT. D–F) Predicted binding sites of CREB1 within the CARD9 promoter region. G) The binding of CREB1 to Card9 promoter in BMDM treated with FMT and BTX via chromatin immunoprecipitation (ChIP) assay. H) Immunofluorescence analysis of CREB1 expression in BMDMs following different treatments. I) Levels of cAMP in BMDMs treated with BTX and FMT. J) Western blot analysis of total PKA (t‐PKA), phosphorylated PKA (p‐PKA), total CREB1 (t‐CREB1), phosphorylated CREB1 (p‐CREB1), and CARD9 in BMDMs under various treatment conditions. K) Nuclear and cytoplasmic expression levels of t‐CREB1 and p‐CREB1 in BMDMs. L) Western blot analysis of t‐PKA, p‐PKA, t‐CREB1, p‐CREB1, and CARD9 expression in BMDMs treated with FMT and KG‐501 (a CREB1 inhibitor). M) Western blot analysis of t‐PKA, p‐PKA, t‐CREB1, p‐CREB1, and CARD9 expression in BMDMs treated with BTX and TTK21 (a CREB1 activator). N) CD206 + macrophage levels in BMDMs treated with FMT and KG‐501. O) CD11c + macrophage levels in BMDMs treated with BTX and TTK21. P) Schematic summary illustrating the proposed mechanism of NE‐mediated regulation of macrophage activation through the PKA/CREB1/CARD9 signaling axis. The data were compiled from at least three independent experiments and are presented as the means ± SEMs. * p < 0.05, ** p < 0.01, *** p < 0.001 by Student's unpaired two‐tailed t ‐test or one‐way analysis of variance (ANOVA) with Tukey's post hoc comparison.

Article Snippet: For β2‐AR studies, cells were pre‐treated with FMT (100 n m ) or BTX (10 μ m ) for 2 h and then treated with NE (100 n m ; HY‐13715, MedChemExpress, USA) for 12 h. [ , , ] The mouse CARD9 expression plasmids and sh Card9 were obtained from Gene Create (Wuhan, China).

Techniques: Activation Assay, Expressing, Binding Assay, Chromatin Immunoprecipitation, Immunofluorescence, Western Blot, Two Tailed Test, Comparison

Journal: Advanced Science

Article Title: CARD9 Conveys Pancreatic Islet Sympathetic Nervous β2 Signals to Reshape Macrophage Creatine Metabolism in Type 1 Diabetes

doi: 10.1002/advs.202507543

Figure Lengend Snippet: β2‐AR signaling mediates macrophage polarization through CARD9‐regulated creatine transport. A) Intracellular ATP levels in macrophages treated with BTX and FMT. B) Schematic diagram illustrating the process of creatine (Cr) metabolism in macrophages. C) Intracellular and extracellular Cr levels in macrophages following treatment with BTX and FMT. D) A scatter plot illustrating the correlations between insulin and Cr in the plasma of diabetes patients. E) Heatmap showing the relative transcription levels of SLC6A8, GAMT, GATM, and CKB in macrophages treated with BTX and FMT. F) Correlation analysis between CARD9 and SLC6A8 expression levels in pancreatic tissues. G) Western blot analysis of SLC6A8 protein expression in BMDMs and RAW264.7 treated with BTX and FMT. H) Flow cytometry analysis of CD11c and CD206 expression under the F4/80 + gating after exogenous administration of Cr and cyclocreatine (CCr) in BMDMs treated with BTX. I) Immunofluorescence analysis of PC12 after co‐culture with exogenous application of Cr and CCr in BMDMs treated with BTX. J) Flow cytometry analysis of CD11c and CD206 expression under the F4/80 + gating after exogenous administration of RGX‐202 (a Cr transporter inhibitor) in BMDMs treated with FMT. K) Flow cytometry analysis of CD11c and CD206 expression under the F4/80 + gating after exogenous administration of Cr and CCr in WT‐BMDMs and Card9 −/− ‐BMDMs. The data were compiled from at least three independent experiments and are presented as the means ± SEMs. * p < 0.05, ** p < 0.01, *** p < 0.001 by Student's unpaired two‐tailed t ‐test or one‐way analysis of variance (ANOVA) with Tukey's post hoc comparison.

Article Snippet: For β2‐AR studies, cells were pre‐treated with FMT (100 n m ) or BTX (10 μ m ) for 2 h and then treated with NE (100 n m ; HY‐13715, MedChemExpress, USA) for 12 h. [ , , ] The mouse CARD9 expression plasmids and sh Card9 were obtained from Gene Create (Wuhan, China).

Techniques: Clinical Proteomics, Expressing, Western Blot, Flow Cytometry, Immunofluorescence, Co-Culture Assay, Two Tailed Test, Comparison