got1 inhibitor  (MedChemExpress)

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: EMBO Reports

Article Title: Glycogenesis and glyconeogenesis from glutamine, lactate and glycerol support human macrophage functions

doi: 10.1038/s44319-024-00278-4

Figure Lengend Snippet: Analysis of the effect of NV-161, CB-839, GOT1i, PEPCKi, MB05032, CHC, GSK2837808, CP-91149, and 6AN on cell viability.

Article Snippet: GOT1i , MedChemExpress , HY-122723.

Techniques: Concentration Assay

Journal: EMBO Reports

Article Title: Glycogenesis and glyconeogenesis from glutamine, lactate and glycerol support human macrophage functions

doi: 10.1038/s44319-024-00278-4

Figure Lengend Snippet: Reagents and tools table

Article Snippet: GOT1i , MedChemExpress , HY-122723.

Techniques: Sequencing, Control, Magnetic Beads, Recombinant, Colorimetric Assay, Staining, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Software, Microscopy, Cytometry, Mass Spectrometry, Imaging, Spectrophotometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: SIRT1 maintains bone homeostasis by regulating osteoblast glycolysis through GOT1

doi: 10.1007/s00018-023-05043-9

Figure Lengend Snippet: SIRT1 inhibited GOT1 enzyme activity by catalyzing its lysine deacetylation. A The concentration of Asp in the femurs of the control and cKO mice aged 8 weeks measured by an ELISA Kit (n = 8/group). B The concentration of malate in the femurs of the control and cKO mice aged 8 weeks measured by an ELISA Kit (n = 8/group). C The activity of GOT1 in the femurs of the control and cKO mice aged 8 weeks measured by an ELISA Kit (n = 8/group). D The subcellular localization of SIRT1 and GOT1 in primary osteoblasts. E The protein level and Kac level of GOT1 upon Sirt1 deletion or overexpression in 3T3E1 cells. F The Kac level of GOT1 in Sirt1 knockout 3T3E1 cells upon Sirt1 overexpression. G The Kac level of GOT1 in 3T3E1 cells treated with SRT2104 or EX-527. H The Kac level of GOT1 protein immunoprecipitated from 3T3E1 cells transfected with SIRT1 plasmids or mutant SIRT1-H355A plasmids. I Control and Sirt1 knockout 3T3E1 cells were transfected with Wild-type GOT1 or GOT1-K318R mutant plasmids. The Kac level of GOT1 protein immunoprecipitated from the above cells. J Sirt1-knockout/Got1- knockout 3T3E1 cells were transfected with Wild-type GOT1 or GOT1-K318R mutant plasmids. The GOT1 activity was measured in the above cells. K Sirt1-knockout/Got1- knockout 3T3E1 cells were transfected with Wild-type GOT1 or GOT1-K318R mutant plasmids. The levels of F6P in glycolysis were measured in the above cells. L Sirt1-knockout/Got1- knockout 3T3E1 cells were transfected with Wild-type GOT1 or GOT1-K318R mutant plasmids. The levels of PEP in glycolysis were measured in the above cells. M Sirt1-knockout/Got1- knockout 3T3E1 cells were infected with Wild-type GOT1 or GOT1-K318R mutant virus. The ALP staining of osteoblast mineralization was conducted in the above cells. Data are presented as the mean ± SD, Student’s t test (a, b, c); one-way ANOVA with Tukey’s test (j, k, l), *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: To inhibit the activity of GOT1, mice aged 8 weeks were treated with vehicle (saline) and/or 10 mg/kg aminooxyacetic acid (AOA) (MedChemExpress, HY-107994) every other day until dissection at 16 weeks, and osteoblasts were treated with vehicle or 100 μM AOA for 48 h.

Techniques: Activity Assay, Concentration Assay, Control, Enzyme-linked Immunosorbent Assay, Over Expression, Knock-Out, Immunoprecipitation, Transfection, Mutagenesis, Infection, Virus, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: SIRT1 maintains bone homeostasis by regulating osteoblast glycolysis through GOT1

doi: 10.1007/s00018-023-05043-9

Figure Lengend Snippet: SIRT1 affected the acetylation level of GOT1 and glycolysis to regulate bone mass. A Schematic of experimental design. The 8-week cKO mice treated with vehicle or AOA until dissection at 16 weeks. B GOT1 activity of 16-week cKO mice treated with vehicle or AOA (n = 8/group). C The concentration of Asp in 16-week cKO mice treated with vehicle or AOA (n = 8/group). D The concentration of malate in 16-week cKO mice treated with vehicle or AOA (n = 8/group). E Representative micro-CT images of the femurs in 16-week cKO mice treated with vehicle or AOA. F Quantification of the BV/TV, Tb.N, Tb.Th, and Tb.Sp from the distal femurs in 16-week cKO mice treated with vehicle or AOA (n = 7/group). G Quantification of the metabolites in glycolysis after AOA treatment in 16-week cKO mice (n = 8/group). H Relative mRNA expression of Runx2, Col1a1, Alpl, and Bglap in Sirt1KO osteoblasts treated with vehicle or AOA (n = 6/group). I Representative images of ALP staining of Sirt1KO osteoblasts treated with vehicle or AOA. J Representative images of Alizarin Red S staining of Sirt1KO osteoblasts treated with vehicle or AOA. K Quantification of caspase-3 activity in Sirt1KO osteoblasts treated with vehicle or AOA (n = 6/group). BV/TV: bone volume fraction; Tb.Th: trabecular thickness; Tb.N: trabecular number; Tb.Sp: trabecular separation; Cort.Th: cortical bone thickness; Tt.Ar: total cortical bone area. Data are presented as the mean ± SD, Student’s t test; *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: To inhibit the activity of GOT1, mice aged 8 weeks were treated with vehicle (saline) and/or 10 mg/kg aminooxyacetic acid (AOA) (MedChemExpress, HY-107994) every other day until dissection at 16 weeks, and osteoblasts were treated with vehicle or 100 μM AOA for 48 h.

Techniques: Dissection, Activity Assay, Concentration Assay, Micro-CT, Expressing, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: SIRT1 maintains bone homeostasis by regulating osteoblast glycolysis through GOT1

doi: 10.1007/s00018-023-05043-9

Figure Lengend Snippet: Schematic diagram of SIRT1 deacetylating GOT1 to regulate glycolysis in osteoblasts. SIRT1 deletion induced acetylation of GOT1, thereby inhibited the process of glycolysis to decrease bone formation by osteoblasts. The decreased bone formation then inhibited bone resorption. Thus, bone mass decreased

Article Snippet: To inhibit the activity of GOT1, mice aged 8 weeks were treated with vehicle (saline) and/or 10 mg/kg aminooxyacetic acid (AOA) (MedChemExpress, HY-107994) every other day until dissection at 16 weeks, and osteoblasts were treated with vehicle or 100 μM AOA for 48 h.

Techniques: