α kg  (Merck & Co)

Journal: Molecular Biomedicine

Article Title: Macrophage Trem2 deficiency aggravates aging-induced vascular remodeling by acting as a non-classical receptor of interleukin-13

doi: 10.1186/s43556-025-00377-1

Figure Lengend Snippet: IL-13/Trem2 combination enhance the beneficial crosstalk between macrophage and VSMCs through Syk-Sp1-SLC25A51 pathway-mediated upregulation of α-KG. a Measuring the α-KG concentration in senescent macrophages upon Ang II stimulation at different time points with/without IL-13 treatment. n = 5 samples/group. b Measuring the α-KG concentration in IL-13-treated senescent macrophages isolated from WT and T2-KO mice. n = 5 samples per group. c Measuring the concentration of α-KG in senescent macrophages treated with IL-13 and then transfected with a Trem2-overexpressing lentivirus. n = 5 samples per group. d Measurement of the α-KG concentration in IL-13-treated T2-KO senescent macrophages transfected a lentivirus for Sp1 or SLC25A51 overexpression. n = 5 samples per group. e Measuring the concentration of α-KG in IL-13-treated T2-KO senescent macrophages transfected a lentivirus for Sirt2, Sirt3, Sirt5, or Nampt overexpression. n = 5 samples/group. f QPCR quantification of contractile protein markers Cnn1, SM-22α, and α-SMA in α-KG-treated senescent VSMCs. n = 6 samples/group. g Measurement of Caspase 3/7 activity in α-KG-treated senescent VSMCs. n = 5 samples per group. h , i DHE staining images and quantification in α-KG-treated senescent VSMCs. Scale bar: 100 µm. n = 5 samples per group. j, k Images and quantification of the wound-healing assay regarding the effects of α-KG on VSMC migration in a co-culture experimental system of IL-13-treated T2-KO senescent macrophages and VSMCs. Scale bar: 100 µm. n = 6 samples per group. l QPCR quantification of the effect of α-KG on contractile protein markers α-SMA in a co-culture experimental system of IL-13-treated T2-KO senescent macrophages and VSMCs. n = 5 samples per group. m, n Immunofluorescence staining and quantification of the effect of α-KG on contractile protein markers α-SMA in a co-culture experimental system of IL-13-treated T2-KO senescent macrophages and VSMCs. Scale bar: 100 µm. n = 5 samples per group. o, p Edu staining images and quantification regarding the effect of α-KG on cell proliferation of VSMCs in a co-culture experimental system of IL-13-treated T2-KO senescent macrophages and VSMCs. Scale bar: 100 µm. n = 5 samples per group. q, r DHE staining images and quantification data regarding the effect of α-KG on cell proliferation of VSMCs in a co-culture experimental system of IL-13-treated T2-KO senescent macrophages and VSMCs. Scale bar: 100 µm. n = 5 samples per group. s, t Caspase 3/7 activity images and quantification data regarding the effect of α-KG on cell apoptosis of VSMCs in a co-culture experimental system of IL-13-treated T2-KO senescent macrophages and VSMCs. Scale bar is 100 µm. n = 5 samples per group. * p < 0.05, ** p < 0.01

Article Snippet: To evaluate the efficacy of α-ketoglutarate (α-KG) supplementation in modulating vascular aging in vivo, 18-month-old male WT and T2-cKO mice were randomly allocated into four experimental groups and provided free access to either standard chow or chow supplemented with 2% α-KG (MedChemExpress, 328–50–7) for six months.

Techniques: Concentration Assay, Isolation, Transfection, Over Expression, Activity Assay, Staining, Wound Healing Assay, Migration, Co-Culture Assay, Immunofluorescence

Journal: Molecular Biomedicine

Article Title: Macrophage Trem2 deficiency aggravates aging-induced vascular remodeling by acting as a non-classical receptor of interleukin-13

doi: 10.1186/s43556-025-00377-1

Figure Lengend Snippet: Administration of α-KG counteracts the vascular ageing and dysfunction triggered by macrophage Trem2 loss in vivo. a PWV measurements in four groups of aged mice. n = 8 samples/group. b Measurement of arterial vessel contractions mediated by PE in four groups of aged mice. n = 8 samples per group. c Measurement of arterial vessel relaxation mediated by SNP in four groups of aged mice. n = 8 samples per group. d H&E staining analysis on aortic samples from four groups of aged mice. Scale bar: 100 µm. n = 8 samples per group. e, f Masson staining images and quantification of aortic samples from aged mice. Scale bar: 100 µm. n = 8 samples per group. g, h DHE staining images and quantification on aortic samples from aged mice. Scale bar: 100 µm. n = 8 samples per group. i-l QPCR quantification of the expressions of Collagen I, α-SMA, IL-6 and Sirt6 in aortic samples from aged mice. n = 5 samples per group. m Working model: The macrophage IL-13/Trem2/Syk-Sp1-SLC25A51 axis mitigates vascular aging by enhancing mitochondrial NAD + transport and α-ketoglutarate (α-KG) production. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: To evaluate the efficacy of α-ketoglutarate (α-KG) supplementation in modulating vascular aging in vivo, 18-month-old male WT and T2-cKO mice were randomly allocated into four experimental groups and provided free access to either standard chow or chow supplemented with 2% α-KG (MedChemExpress, 328–50–7) for six months.

Techniques: In Vivo, Staining

Journal: Experimental & Molecular Medicine

Article Title: SLC6A14-mediated glutamine promotes SYTL4–CXCL8 axis activation to drive gemcitabine resistance and immune evasion in pancreatic cancer

doi: 10.1038/s12276-025-01596-w

Figure Lengend Snippet: a The peak area of α-KG in CAPAN-1, CAPAN-1/GR si Con and CAPAN-1/GR si SLC6A14 incubated for 24 h with a medium containing ±glutamine was measured by LC–MS analysis. b Intracellular α-KG levels in CAPAN-1, CAPAN-1/GR si Con and CAPAN-1/GR si SLC6A14 incubated for 24 h with a medium containing ±glutamine, followed by glutamine assays. c Western blot analysis of SLC6A14, p-mTOR, mTOR, p-NF-κB and NF-κB in CAPAN-1, CAPAN-1/GR si Con and CAPAN-1/GR si SLC6A14 incubated for 24 h with ±glutamine. d Cytokine expression profiles in CAPAN-1/GR si Con versus si SLC6A14 via cytokine assays. Cytokine detection (top left), heatmap (bottom left) and P values (right). e CXCL8 levels in CAPAN-1, CAPAN-1/GR si Con , CAPAN-1/GR si SLC6A14 or α-MT for 24 h, then incubated with ±glutamine for 24 h. f GSEA of downregulated genes involved in the positive regulation of secretion in CAPAN-1/GR cells si Con versus si SLC6A14 (FDR q value <0.05; top). KEGG enrichment analysis of genes downregulated in CAPAN-1/GR cells transfected with si SLC6A14 (bottom). g A set of regulated secretion, exocytosis and SLC6A14 -related genes were compared, and two common genes were identified. Correlation analyses of CXCL8 between either ANXA2 or SYTL4 were performed using TISIDB. h Kaplan–Meier survival analysis of patients with PDAC based on SLC6A14 and SYTL4 expression for overall survival graph. i Endogenous SYTL4 was immunoprecipitated with endogenous SLC6A14 and NF-κB from CAPAN-1/GR cells and subjected to immunoprecipitation, followed by immunoblotting with indicated antibodies. j CAPAN-1/GR cells si SLC6A14 were subjected to immunoprecipitation, followed by immunoblot analysis with indicated antibodies. k CXCL8 secretion levels in CAPAN-1, CAPAN-1/GR si Con and CAPAN-1/GR si SLC6A14 or si SYTL4 , and CAPAN-1/GR cells cotransfected with both si SLC6A14 and si SYTL4 . Error bars, mean ± s.d., # P < 0.05; ## P < 0.01; ### P < 0.001; n.s., not significant; by Student’s t -test ( a , b , e and h ).

Article Snippet: For qualitative analysis, α-ketoglutarate (α-KG) analytical standard (Merck) was manufactured by concentration with MeOH, mixed with ISTD solution and then measured as standards for LC–MS analysis.

Techniques: Incubation, Liquid Chromatography with Mass Spectroscopy, Western Blot, Expressing, Transfection, Immunoprecipitation