anti got1  (Proteintech)

Journal: Advanced Science

Article Title: GOT1 Inhibition Induces Extracellular Matrix Remodeling in Pancreatic Cancer

doi: 10.1002/advs.202516578

Figure Lengend Snippet: Effects of GOT1 inhibition in the pancreatic TME. (a) Schematic of glutamine metabolism. (b) Determination of half‐maximal inhibitory concentration (IC 50 ) of the GOT1 inhibitor using mono‐ and multicellular 3D cultures. (c) Cell viability of mono‐ and multicellular 3D cultures. n = 3, **** = p ≤ 0.0001, (d) live/dead staining (scale bar = 100 µm), and (e) relative metabolite quantification following GOT1 inhibition. (f) Gene ontology enrichment analysis, and (g) volcano plot showing the significantly up/downregulated matrisome proteins of treated versus non‐treated multicellular 3D cultures. Blue = upregulated, red = downregulated. FDR cut‐off = 0.05, log2(fold‐change) ≥ 2. (h) Immunoblot of multicellular 3D cultures upon GOT1 inhibition. (i) Cell metabolic activity for PDAC cells cultured in type III collagen. n = 6, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001. (j) Cytokine profiling of multicellular 3D cultures following GOT1 inhibition. GOT1, glutamic‐oxaloacetic transaminase 1. TME, tumor microenvironment. PDAC, pancreatic ductal adenocarcinoma.

Article Snippet: Mono‐ and multicellular 3D cultures were established as described above and grown for 7 days and then treated with 25 μM GOT1 inhibitor (MedchemExpress HY‐122723) alone or combined with 100 nM gemcitabine (Sigma‐Aldrich G6423) and 100 nM nab‐paclitaxel (Bristol‐Myers Squibb), which were dissolved directly into PDAC or TKCC cell media.

Techniques: Inhibition, Concentration Assay, Staining, Western Blot, Activity Assay, Cell Culture

Journal: Advanced Science

Article Title: GOT1 Inhibition Induces Extracellular Matrix Remodeling in Pancreatic Cancer

doi: 10.1002/advs.202516578

Figure Lengend Snippet: Analyses of cell responses following combined GOT1 inhibition and treatment with cytotoxic drugs. (a) Schematic of experimental rationale. (b) Cell viability of mono‐ and multicellular 3D cultures grown in GelMA‐HAMA hydrogels following treatment with GOT1 inhibitor ± gemcitabine and nab‐paclitaxel. n = 3, * = p ≤ 0.05, *** = p ≤ 0.001. (c) Gene ontology enrichment analysis, (d) signaling pathway analysis, and (e) volcano plot showing significantly up/downregulated matrisome proteins of treated multicellular 3D cultures compared to non‐treated cells grown in GelMA‐HAMA hydrogels. Blue = upregulated, red = downregulated. (f) Relative metabolite quantification in multicellular 3D cultures following treatment with GOT1 inhibitor ± gemcitabine and nab‐paclitaxel. FDR cut‐off = 0.05, log2(fold‐change) ≥ 2. GOT1, glutamic‐oxaloacetic transaminase 1. GelMA, gelatin methacrylate. HAMA, hyaluronic acid methacrylate.

Article Snippet: Mono‐ and multicellular 3D cultures were established as described above and grown for 7 days and then treated with 25 μM GOT1 inhibitor (MedchemExpress HY‐122723) alone or combined with 100 nM gemcitabine (Sigma‐Aldrich G6423) and 100 nM nab‐paclitaxel (Bristol‐Myers Squibb), which were dissolved directly into PDAC or TKCC cell media.

Techniques: Inhibition

Journal: Advanced Science

Article Title: GOT1 Inhibition Induces Extracellular Matrix Remodeling in Pancreatic Cancer

doi: 10.1002/advs.202516578

Figure Lengend Snippet: Multicellular 3D cultures including patient‐derived PDAC cells and their response toward GOT1 inhibition. (a) Micrograph of multicellular 3D cultures grown in GelMA‐HAMA hydrogels (scale bar = 100 µm). (b) Cell metabolic activity and (c) DNA content of mono‐ and multicellular 3D cultures including patient‐derived PDAC cells in GelMA‐HAMA hydrogels. (d) Cell viability, and (e) DNA content of mono‐ and multicellular TKCC‐22‐LO cultures following treatment with GOT1 inhibitor ± gemcitabine and nab‐paclitaxel. n = 3, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001, **** = p ≤ 0.0001. (f) Live/dead staining (scale bar = 100 µm), and (g) heatmap of amino acid metabolism microarray of multicellular 3D cultures treated with the GOT1 inhibitor compared to non‐treated cells. PDAC, pancreatic ductal adenocarcinoma. GOT1, glutamic‐oxaloacetic transaminase 1. GelMA, gelatin methacrylate. HAMA, hyaluronic acid methacrylate.

Article Snippet: Mono‐ and multicellular 3D cultures were established as described above and grown for 7 days and then treated with 25 μM GOT1 inhibitor (MedchemExpress HY‐122723) alone or combined with 100 nM gemcitabine (Sigma‐Aldrich G6423) and 100 nM nab‐paclitaxel (Bristol‐Myers Squibb), which were dissolved directly into PDAC or TKCC cell media.

Techniques: Derivative Assay, Inhibition, Activity Assay, Staining, Microarray

Journal: Journal of Translational Medicine

Article Title: FFAR4 negatively regulates colorectal cancer growth via blocking oxidative phosphorylation

doi: 10.1186/s12967-026-07942-4

Figure Lengend Snippet: TUG891 reduces the NAD⁺/NADH ratio and disrupts mitochondrial redox homeostasis. ( A - B ) Principal-component analysis (PCA) of gene expression data, showing the clustering of gene profiles for control and FFAR4 activation groups in HCT116 cells ( A ) and HT29 cells ( B ). ( C ) Changes in metabolite levels with and without TUG891 treatment in HCT116 cells (upper panel) and HT29 cells (lower panel). Log2(FC) indicates log2-transformed fold change of the TUG891-treated group relative to the control group. ( D - E ) Over-Representation Analysis identified significantly enriched KEGG pathways based on different metabolites in HCT116 cells ( D ) and HT29 cells ( E ). ( F ) Western blot analysis of malate–aspartate shuttle–related enzymes (GOT1, GOT2, MDH1, and MDH2) and OXPHOS proteins in control and TUG891-treated cells. ( G - H ) Measurement of NAD + levels and the NAD + /NADH ratio in HCT116 cells comparing vehicle and FFAR4 activator groups. ( I - J ) Measurement of NAD + levels and the NAD + /NADH ratio in HT29 cells comparing vehicle and FFAR4 activator groups. ( K , L ) Measurement of the cellular ATP/ADP ratio in HCT116 ( K ) and HT29 ( L ) cells comparing vehicle and FFAR4 activator–treated groups. The data were expressed as the mean ± standard deviation (SD)

Article Snippet: The primary antibodies included Cyclin D1 (Proteintech, China), GOT1 (ABclonal, China), GOT2 (ABclonal, China), MDH1 (ABclonal, China), MDH2 (ABclonal, China), OXPHOS Cocktail (Proteintech, China), and β-actin (Cell Signaling Technology, USA), all diluted 1:1000 in TBST containing 3% bovine serum albumin (BSA).

Techniques: Gene Expression, Control, Activation Assay, Transformation Assay, Western Blot, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: FFAR4 negatively regulates colorectal cancer growth via blocking oxidative phosphorylation

doi: 10.1186/s12967-026-07942-4

Figure Lengend Snippet: TUG891 suppresses tumor growth in an MC38 syngeneic mouse model. ( A ) Body weight of mice treated with PBS or TUG891 during the experimental period ( n = 7). The data were expressed as the mean ± standard deviation (SD). ( B ) Tumor growth curves showing tumor volume over time ( n = 7). The data were expressed as the mean ± Error of the Mean (SEM). ( C ) Representative images of excised tumors at the experimental endpoint ( n = 7). ( D ) Tumor weights measured at sacrifice ( n = 7). The data were expressed as the mean ± standard deviation (SD). ( E ) Immunofluorescence staining of FFAR4 in tumor tissues from PBS- and TUG891-treated mice. ( F ) Western blot analysis of GOT1, MDH1, and oxidative phosphorylation–related proteins (ATP5A1 and UQCRC1) in tumor tissues. ns, not significant ( P ≥ 0.05)

Article Snippet: The primary antibodies included Cyclin D1 (Proteintech, China), GOT1 (ABclonal, China), GOT2 (ABclonal, China), MDH1 (ABclonal, China), MDH2 (ABclonal, China), OXPHOS Cocktail (Proteintech, China), and β-actin (Cell Signaling Technology, USA), all diluted 1:1000 in TBST containing 3% bovine serum albumin (BSA).

Techniques: Standard Deviation, Immunofluorescence, Staining, Western Blot, Phospho-proteomics