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  • 86
    BOC Sciences 13699-48-4
    13699 48 4, supplied by BOC Sciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/13699-48-4/product/BOC Sciences
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    13699-48-4 - by Bioz Stars, 2024-02
    86/100 stars
      Buy from Supplier

    92
    Proteintech fmo5
    C. elegans FMO-2 and mammalian FMOs improve stress resistance to oxidative stress. (A) Amino acid sequence alignment of FMOs across species with the regions containing the eight essential residues denoted as red arrowheads in the catalytic active site among reconstructed ancestral mammalian <t>FMO5,</t> C. elegans FMO-2, and mouse <t>FMO1–5.</t> (B) Wild-Type and FMO-2 overexpressing (FMO-2 OE) worm survival curves on paraquat stress. Each strain was placed on NGM (Nematode Growth Medium) containing 5 mM paraquat from the fourth larvae stage (L4) at Day 0. Survival was quantified every day until all worms were dead. The difference between the survival curves is denoted with the log-rank test p -value. (C) FMO1–5 protein levels in FMO1–5 OE and control cells of both HEK293A and HepG2. (D) FMO5 OE and control cell survival curves on paraquat stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing doses of paraquat. (E) LD 50 values of FMO1–5 OE cells compared to the control cells on paraquat stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO1–5 or empty vector pDEST were subjected to indicated increasing doses of paraquat. (F) FMO5 OE and control cell survival curves on cadmium stress in HEK 293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing doses of cadmium. (G) LD 50 values of FMO-OE cells compared to the control cells on cadmium stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO or empty vector pDEST were subjected to indicated increasing doses of paraquat. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    Fmo5, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fmo5/product/Proteintech
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fmo5 - by Bioz Stars, 2024-02
    92/100 stars
      Buy from Supplier

    92
    Proteintech 13699 1 ap
    C. elegans FMO-2 and mammalian FMOs improve stress resistance to oxidative stress. (A) Amino acid sequence alignment of FMOs across species with the regions containing the eight essential residues denoted as red arrowheads in the catalytic active site among reconstructed ancestral mammalian <t>FMO5,</t> C. elegans FMO-2, and mouse <t>FMO1–5.</t> (B) Wild-Type and FMO-2 overexpressing (FMO-2 OE) worm survival curves on paraquat stress. Each strain was placed on NGM (Nematode Growth Medium) containing 5 mM paraquat from the fourth larvae stage (L4) at Day 0. Survival was quantified every day until all worms were dead. The difference between the survival curves is denoted with the log-rank test p -value. (C) FMO1–5 protein levels in FMO1–5 OE and control cells of both HEK293A and HepG2. (D) FMO5 OE and control cell survival curves on paraquat stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing doses of paraquat. (E) LD 50 values of FMO1–5 OE cells compared to the control cells on paraquat stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO1–5 or empty vector pDEST were subjected to indicated increasing doses of paraquat. (F) FMO5 OE and control cell survival curves on cadmium stress in HEK 293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing doses of cadmium. (G) LD 50 values of FMO-OE cells compared to the control cells on cadmium stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO or empty vector pDEST were subjected to indicated increasing doses of paraquat. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    13699 1 Ap, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/13699 1 ap/product/Proteintech
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    13699 1 ap - by Bioz Stars, 2024-02
    92/100 stars
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    92
    Proteintech rabbit anti fmo5 polyclonal antibody
    Gene targeting and generation of <t>Fmo5</t> −/− KO mice. (A) Recombination between homologous regions (thick black lines) of the targeting construct and the Fmo5 locus leading to the generation of the Fmo5 targeted allele, in which exon 2 is replaced by the neo r cassette (neo). Exons are represented by black boxes. Primers used in PCR screening of the G418 r ES clones are depicted with black arrows (a and b). Primers used to distinguish between the WT and targeted alleles are depicted with white arrows (c and d) (see also C). Restriction sites and probes used for Southern blot genotyping of the targeted ES clones and the mice are also shown. Not drawn to scale. (B) After PCR screening (not shown) Southern blot analysis with (i) a neo r -specific probe, (ii) a 5′-internal probe, (iii) a 3′-external probe further confirmed the correct structure of the targeted allele. Analysis using a vector-specific probe verified the absence of any random integrations in the targeted ES clones (data not shown). (C) Mice were genotyped by PCR analysis of tail DNA with primers c and d (see A). Het, heterozygous knockout ( Fmo5 +/− ); KO, homozygous knockout ( Fmo5 −/− ); WT, wild type. (D) Western blot of liver proteins showing lack of FMO5 expression in Fmo5 −/− mice (KO) compared with WT mice (WT).
    Rabbit Anti Fmo5 Polyclonal Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti fmo5 polyclonal antibody/product/Proteintech
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti fmo5 polyclonal antibody - by Bioz Stars, 2024-02
    92/100 stars
      Buy from Supplier

    N/A
    Standard format Plasmid sent in bacteria as agar stab
      Buy from Supplier

    Image Search Results


    C. elegans FMO-2 and mammalian FMOs improve stress resistance to oxidative stress. (A) Amino acid sequence alignment of FMOs across species with the regions containing the eight essential residues denoted as red arrowheads in the catalytic active site among reconstructed ancestral mammalian FMO5, C. elegans FMO-2, and mouse FMO1–5. (B) Wild-Type and FMO-2 overexpressing (FMO-2 OE) worm survival curves on paraquat stress. Each strain was placed on NGM (Nematode Growth Medium) containing 5 mM paraquat from the fourth larvae stage (L4) at Day 0. Survival was quantified every day until all worms were dead. The difference between the survival curves is denoted with the log-rank test p -value. (C) FMO1–5 protein levels in FMO1–5 OE and control cells of both HEK293A and HepG2. (D) FMO5 OE and control cell survival curves on paraquat stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing doses of paraquat. (E) LD 50 values of FMO1–5 OE cells compared to the control cells on paraquat stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO1–5 or empty vector pDEST were subjected to indicated increasing doses of paraquat. (F) FMO5 OE and control cell survival curves on cadmium stress in HEK 293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing doses of cadmium. (G) LD 50 values of FMO-OE cells compared to the control cells on cadmium stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO or empty vector pDEST were subjected to indicated increasing doses of paraquat. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Flavin-Containing Monooxygenases Are Conserved Regulators of Stress Resistance and Metabolism

    doi: 10.3389/fcell.2021.630188

    Figure Lengend Snippet: C. elegans FMO-2 and mammalian FMOs improve stress resistance to oxidative stress. (A) Amino acid sequence alignment of FMOs across species with the regions containing the eight essential residues denoted as red arrowheads in the catalytic active site among reconstructed ancestral mammalian FMO5, C. elegans FMO-2, and mouse FMO1–5. (B) Wild-Type and FMO-2 overexpressing (FMO-2 OE) worm survival curves on paraquat stress. Each strain was placed on NGM (Nematode Growth Medium) containing 5 mM paraquat from the fourth larvae stage (L4) at Day 0. Survival was quantified every day until all worms were dead. The difference between the survival curves is denoted with the log-rank test p -value. (C) FMO1–5 protein levels in FMO1–5 OE and control cells of both HEK293A and HepG2. (D) FMO5 OE and control cell survival curves on paraquat stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing doses of paraquat. (E) LD 50 values of FMO1–5 OE cells compared to the control cells on paraquat stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO1–5 or empty vector pDEST were subjected to indicated increasing doses of paraquat. (F) FMO5 OE and control cell survival curves on cadmium stress in HEK 293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing doses of cadmium. (G) LD 50 values of FMO-OE cells compared to the control cells on cadmium stress in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO or empty vector pDEST were subjected to indicated increasing doses of paraquat. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The following antibodies were used: mouse monoclonal antibody against FMO2 (Proteintech 67019-1-Ig) and polyclonal antibodies against FMO1 (Invitrogen PA5-95285), FMO3 (Abcam ab126711), FMO4 (Invitrogen PA5-79276), FMO5 (Proteintech 13699-1-AP), JNKs (Cell Signaling Technology 9252), phospho-SAPK/JNK (Thr183/Tyr185) (Cell Signaling Technology 9251), ERK1/2 (Cell Signaling Technology 4695), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology 9101), p38 (Cell Signaling Technology 9212), phospho-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology 9211), and GAPDH (Cell Signaling Technology 2118).

    Techniques: Sequencing, Stable Transfection, Expressing, Plasmid Preparation

    FMOs improve stress resistance to broader stressors in mammalian cells. (A,B) FMO1–5 OE and control cell survival curves on arsenite stress in HEK293A and HepG2. HEK293A cells (A) or HepG2 cells (B) stably expressing FMO1–5 or empty vector pDEST were subjected to indicated increasing doses of arsenite. (C) FMO5 OE and control cell survival curves on UV-radiation in HEK 293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing energies of UV-radiation. (D) LD 50 values of FMO1–5 OE cells compared to the control cells on UV-radiation in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO1–5 or empty vector pDEST were subjected to indicated increasing energies of UV-radiation. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Flavin-Containing Monooxygenases Are Conserved Regulators of Stress Resistance and Metabolism

    doi: 10.3389/fcell.2021.630188

    Figure Lengend Snippet: FMOs improve stress resistance to broader stressors in mammalian cells. (A,B) FMO1–5 OE and control cell survival curves on arsenite stress in HEK293A and HepG2. HEK293A cells (A) or HepG2 cells (B) stably expressing FMO1–5 or empty vector pDEST were subjected to indicated increasing doses of arsenite. (C) FMO5 OE and control cell survival curves on UV-radiation in HEK 293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO5 or empty vector pDEST were subjected to indicated increasing energies of UV-radiation. (D) LD 50 values of FMO1–5 OE cells compared to the control cells on UV-radiation in HEK293A and HepG2. HEK293A cells or HepG2 cells stably expressing FMO1–5 or empty vector pDEST were subjected to indicated increasing energies of UV-radiation. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The following antibodies were used: mouse monoclonal antibody against FMO2 (Proteintech 67019-1-Ig) and polyclonal antibodies against FMO1 (Invitrogen PA5-95285), FMO3 (Abcam ab126711), FMO4 (Invitrogen PA5-79276), FMO5 (Proteintech 13699-1-AP), JNKs (Cell Signaling Technology 9252), phospho-SAPK/JNK (Thr183/Tyr185) (Cell Signaling Technology 9251), ERK1/2 (Cell Signaling Technology 4695), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology 9101), p38 (Cell Signaling Technology 9212), phospho-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology 9211), and GAPDH (Cell Signaling Technology 2118).

    Techniques: Stable Transfection, Expressing, Plasmid Preparation

    JNK kinase activity is increased in FMO-overexpressing cells under cadmium-induced oxidative stress. (A) SAPK levels and phosphorylation including JNK, p38, and ERK after 10 μM cadmium treatment of indicated time. HEK293A cells were treated with cadmium, and the SAPK activities were measured with antibodies against phosphorylated SAPKs. (B) SAPKs levels and phosphorylation including JNK, p38, and ERK in FMO1–5 OE HEK293A cells and empty vector control cells after 10 μM Cadmium treatment for 4 h. Quantitation of the phosphorylated JNK (C) , p38 (D) , and ERK (E) after 10 μM Cadmium treatment for 4 h in FMO 1–5 OE HEK293A cells [lane 8–12 in (B) ] compared to empty vector control HEK293A cells [lane 7 in (B) ]. (F) SAPK levels and phosphorylation including JNK, p38, and ERK in FMO1–5 OE HepG2 cells and empty vector control cells after 10 μM Cadmium treatment for 4 h. Quantitation of the phosphorylated JNK (G) , p38 (H) , and ERK (I) after 10 μM Cadmium treatment for 4 h in FMO1–5 OE HepG2 cells [lane 8–12 in (F) ] compared to empty vector control HepG2 cells [lane 7 in (F) ]. Western blot band intensities were quantified by Image J. The phosphorylated bands were normalized to the corresponding unphosphorylated bands of each SAPK, and then were compared to the empty vector pDEST control as fold changes. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Flavin-Containing Monooxygenases Are Conserved Regulators of Stress Resistance and Metabolism

    doi: 10.3389/fcell.2021.630188

    Figure Lengend Snippet: JNK kinase activity is increased in FMO-overexpressing cells under cadmium-induced oxidative stress. (A) SAPK levels and phosphorylation including JNK, p38, and ERK after 10 μM cadmium treatment of indicated time. HEK293A cells were treated with cadmium, and the SAPK activities were measured with antibodies against phosphorylated SAPKs. (B) SAPKs levels and phosphorylation including JNK, p38, and ERK in FMO1–5 OE HEK293A cells and empty vector control cells after 10 μM Cadmium treatment for 4 h. Quantitation of the phosphorylated JNK (C) , p38 (D) , and ERK (E) after 10 μM Cadmium treatment for 4 h in FMO 1–5 OE HEK293A cells [lane 8–12 in (B) ] compared to empty vector control HEK293A cells [lane 7 in (B) ]. (F) SAPK levels and phosphorylation including JNK, p38, and ERK in FMO1–5 OE HepG2 cells and empty vector control cells after 10 μM Cadmium treatment for 4 h. Quantitation of the phosphorylated JNK (G) , p38 (H) , and ERK (I) after 10 μM Cadmium treatment for 4 h in FMO1–5 OE HepG2 cells [lane 8–12 in (F) ] compared to empty vector control HepG2 cells [lane 7 in (F) ]. Western blot band intensities were quantified by Image J. The phosphorylated bands were normalized to the corresponding unphosphorylated bands of each SAPK, and then were compared to the empty vector pDEST control as fold changes. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The following antibodies were used: mouse monoclonal antibody against FMO2 (Proteintech 67019-1-Ig) and polyclonal antibodies against FMO1 (Invitrogen PA5-95285), FMO3 (Abcam ab126711), FMO4 (Invitrogen PA5-79276), FMO5 (Proteintech 13699-1-AP), JNKs (Cell Signaling Technology 9252), phospho-SAPK/JNK (Thr183/Tyr185) (Cell Signaling Technology 9251), ERK1/2 (Cell Signaling Technology 4695), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology 9101), p38 (Cell Signaling Technology 9212), phospho-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology 9211), and GAPDH (Cell Signaling Technology 2118).

    Techniques: Activity Assay, Plasmid Preparation, Quantitation Assay, Western Blot

    FMO expression increases mitochondrial respiration. (A) Mitochondrial respiration measured by OCR (oxygen consumption rate) in FMO1–5 OE HEK293A cells and empty vector control cells. Mitochondrial respiration chain complex inhibitors Oligomycin, Carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP), and rotenone/antimycin A were injected stepwise as indicated. As denoted in the first panel, basal respiration, ATP production, maximal respiration, and spare respiration can be calculated by the OCR level changes in response to the inhibitor injections. (B) Basal respiration, ATP production, maximal respiration, and spare capacity in FMO1–5 OE HEK293A cells and empty vector control cells. (C) Mitochondrial respiration in FMO1–5 OE HepG2 cells. (D) Basal respiration, ATP production, maximal respiration, and spare capacity in FMO1–5 OE HepG2 cells. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Flavin-Containing Monooxygenases Are Conserved Regulators of Stress Resistance and Metabolism

    doi: 10.3389/fcell.2021.630188

    Figure Lengend Snippet: FMO expression increases mitochondrial respiration. (A) Mitochondrial respiration measured by OCR (oxygen consumption rate) in FMO1–5 OE HEK293A cells and empty vector control cells. Mitochondrial respiration chain complex inhibitors Oligomycin, Carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP), and rotenone/antimycin A were injected stepwise as indicated. As denoted in the first panel, basal respiration, ATP production, maximal respiration, and spare respiration can be calculated by the OCR level changes in response to the inhibitor injections. (B) Basal respiration, ATP production, maximal respiration, and spare capacity in FMO1–5 OE HEK293A cells and empty vector control cells. (C) Mitochondrial respiration in FMO1–5 OE HepG2 cells. (D) Basal respiration, ATP production, maximal respiration, and spare capacity in FMO1–5 OE HepG2 cells. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The following antibodies were used: mouse monoclonal antibody against FMO2 (Proteintech 67019-1-Ig) and polyclonal antibodies against FMO1 (Invitrogen PA5-95285), FMO3 (Abcam ab126711), FMO4 (Invitrogen PA5-79276), FMO5 (Proteintech 13699-1-AP), JNKs (Cell Signaling Technology 9252), phospho-SAPK/JNK (Thr183/Tyr185) (Cell Signaling Technology 9251), ERK1/2 (Cell Signaling Technology 4695), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology 9101), p38 (Cell Signaling Technology 9212), phospho-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology 9211), and GAPDH (Cell Signaling Technology 2118).

    Techniques: Expressing, Plasmid Preparation, Injection

    FMO overexpression decreases overall glycolytic activity. (A) Overall glycolytic activity measured by ECAR (extracellular acidification rates) in HEK293A FMO1–5 OE cells and empty vector control cells. Sequential injections of glucose, oligomycin, and 2-DG were applied over time as indicated. As denoted in the first panel, glycolysis, glycolytic capacity, and glycolytic reserve can be calculated by the ECAR level changes in response to individual injections. (B) Glycolysis, glycolytic capacity, and glycolytic reserve in HEK293A FMO1–5 OE cells and empty vector control cells. (C) Overall glycolytic activity in FMO1–5 OE HepG2 cells. (D) Glycolysis, glycolytic capacity, and glycolytic reserve in FMO1–5 OE HepG2 cells and empty vector control cells. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Flavin-Containing Monooxygenases Are Conserved Regulators of Stress Resistance and Metabolism

    doi: 10.3389/fcell.2021.630188

    Figure Lengend Snippet: FMO overexpression decreases overall glycolytic activity. (A) Overall glycolytic activity measured by ECAR (extracellular acidification rates) in HEK293A FMO1–5 OE cells and empty vector control cells. Sequential injections of glucose, oligomycin, and 2-DG were applied over time as indicated. As denoted in the first panel, glycolysis, glycolytic capacity, and glycolytic reserve can be calculated by the ECAR level changes in response to individual injections. (B) Glycolysis, glycolytic capacity, and glycolytic reserve in HEK293A FMO1–5 OE cells and empty vector control cells. (C) Overall glycolytic activity in FMO1–5 OE HepG2 cells. (D) Glycolysis, glycolytic capacity, and glycolytic reserve in FMO1–5 OE HepG2 cells and empty vector control cells. Data represent mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The following antibodies were used: mouse monoclonal antibody against FMO2 (Proteintech 67019-1-Ig) and polyclonal antibodies against FMO1 (Invitrogen PA5-95285), FMO3 (Abcam ab126711), FMO4 (Invitrogen PA5-79276), FMO5 (Proteintech 13699-1-AP), JNKs (Cell Signaling Technology 9252), phospho-SAPK/JNK (Thr183/Tyr185) (Cell Signaling Technology 9251), ERK1/2 (Cell Signaling Technology 4695), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology 9101), p38 (Cell Signaling Technology 9212), phospho-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology 9211), and GAPDH (Cell Signaling Technology 2118).

    Techniques: Over Expression, Activity Assay, Plasmid Preparation

    FMOs regulate amino acid and energetic metabolic pathways. Untargeted metabolomics in FMO1–5 OE cells ( A , see also – ) compared to empty vector control cells, and the significantly regulated metabolic pathways that are enriched. The abundance analyses of metabolites in central carbon metabolism in FMO1–5 OE cells compared to empty vector control cells ( B,C , see also – ). (A) Metabolic pathways regulated by FMO1 are plotted by the enrichment factor (obtained by dividing “significant hits” by “expected hits” for each pathway) on the x -axis and –log of the p -value on the y -axis. Red indicates significantly changed pathways with p < 0.001. Shared significantly regulated metabolic pathways by more than three FMOs in FMO1–5 are indicated in bold text, including amino acid metabolism (Glycine, serine, and threonine metabolism; Cyanoamino acid metabolism; Aminoacyl-tRNA biosynthesis; Selenoamino acid metabolism; Taurine and hypotaurine metabolism; Cysteine and methionine metabolism; Arginine and proline metabolism; Lysine biosynthesis; Alanine, aspartate, and glutamate metabolism) and metabolism of cofactors and vitamins (Pantothenate and CoA biosynthesis; Vitamin B6 metabolism) (see also – ). (B) The levels of the top 25 changed metabolites between FMO1-OE cells and empty vector control cells are shown in heat map. (C) The levels of metabolites significantly regulated by FMO1 are shown in FMO1-OE cells and empty vector control cells.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Flavin-Containing Monooxygenases Are Conserved Regulators of Stress Resistance and Metabolism

    doi: 10.3389/fcell.2021.630188

    Figure Lengend Snippet: FMOs regulate amino acid and energetic metabolic pathways. Untargeted metabolomics in FMO1–5 OE cells ( A , see also – ) compared to empty vector control cells, and the significantly regulated metabolic pathways that are enriched. The abundance analyses of metabolites in central carbon metabolism in FMO1–5 OE cells compared to empty vector control cells ( B,C , see also – ). (A) Metabolic pathways regulated by FMO1 are plotted by the enrichment factor (obtained by dividing “significant hits” by “expected hits” for each pathway) on the x -axis and –log of the p -value on the y -axis. Red indicates significantly changed pathways with p < 0.001. Shared significantly regulated metabolic pathways by more than three FMOs in FMO1–5 are indicated in bold text, including amino acid metabolism (Glycine, serine, and threonine metabolism; Cyanoamino acid metabolism; Aminoacyl-tRNA biosynthesis; Selenoamino acid metabolism; Taurine and hypotaurine metabolism; Cysteine and methionine metabolism; Arginine and proline metabolism; Lysine biosynthesis; Alanine, aspartate, and glutamate metabolism) and metabolism of cofactors and vitamins (Pantothenate and CoA biosynthesis; Vitamin B6 metabolism) (see also – ). (B) The levels of the top 25 changed metabolites between FMO1-OE cells and empty vector control cells are shown in heat map. (C) The levels of metabolites significantly regulated by FMO1 are shown in FMO1-OE cells and empty vector control cells.

    Article Snippet: The following antibodies were used: mouse monoclonal antibody against FMO2 (Proteintech 67019-1-Ig) and polyclonal antibodies against FMO1 (Invitrogen PA5-95285), FMO3 (Abcam ab126711), FMO4 (Invitrogen PA5-79276), FMO5 (Proteintech 13699-1-AP), JNKs (Cell Signaling Technology 9252), phospho-SAPK/JNK (Thr183/Tyr185) (Cell Signaling Technology 9251), ERK1/2 (Cell Signaling Technology 4695), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology 9101), p38 (Cell Signaling Technology 9212), phospho-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology 9211), and GAPDH (Cell Signaling Technology 2118).

    Techniques: Plasmid Preparation

    Gene targeting and generation of Fmo5 −/− KO mice. (A) Recombination between homologous regions (thick black lines) of the targeting construct and the Fmo5 locus leading to the generation of the Fmo5 targeted allele, in which exon 2 is replaced by the neo r cassette (neo). Exons are represented by black boxes. Primers used in PCR screening of the G418 r ES clones are depicted with black arrows (a and b). Primers used to distinguish between the WT and targeted alleles are depicted with white arrows (c and d) (see also C). Restriction sites and probes used for Southern blot genotyping of the targeted ES clones and the mice are also shown. Not drawn to scale. (B) After PCR screening (not shown) Southern blot analysis with (i) a neo r -specific probe, (ii) a 5′-internal probe, (iii) a 3′-external probe further confirmed the correct structure of the targeted allele. Analysis using a vector-specific probe verified the absence of any random integrations in the targeted ES clones (data not shown). (C) Mice were genotyped by PCR analysis of tail DNA with primers c and d (see A). Het, heterozygous knockout ( Fmo5 +/− ); KO, homozygous knockout ( Fmo5 −/− ); WT, wild type. (D) Western blot of liver proteins showing lack of FMO5 expression in Fmo5 −/− mice (KO) compared with WT mice (WT).

    Journal: Biochemical Pharmacology

    Article Title: The phenotype of a knockout mouse identifies flavin-containing monooxygenase 5 (FMO5) as a regulator of metabolic ageing

    doi: 10.1016/j.bcp.2015.05.013

    Figure Lengend Snippet: Gene targeting and generation of Fmo5 −/− KO mice. (A) Recombination between homologous regions (thick black lines) of the targeting construct and the Fmo5 locus leading to the generation of the Fmo5 targeted allele, in which exon 2 is replaced by the neo r cassette (neo). Exons are represented by black boxes. Primers used in PCR screening of the G418 r ES clones are depicted with black arrows (a and b). Primers used to distinguish between the WT and targeted alleles are depicted with white arrows (c and d) (see also C). Restriction sites and probes used for Southern blot genotyping of the targeted ES clones and the mice are also shown. Not drawn to scale. (B) After PCR screening (not shown) Southern blot analysis with (i) a neo r -specific probe, (ii) a 5′-internal probe, (iii) a 3′-external probe further confirmed the correct structure of the targeted allele. Analysis using a vector-specific probe verified the absence of any random integrations in the targeted ES clones (data not shown). (C) Mice were genotyped by PCR analysis of tail DNA with primers c and d (see A). Het, heterozygous knockout ( Fmo5 +/− ); KO, homozygous knockout ( Fmo5 −/− ); WT, wild type. (D) Western blot of liver proteins showing lack of FMO5 expression in Fmo5 −/− mice (KO) compared with WT mice (WT).

    Article Snippet: For FMO5, the blot was incubated with Rabbit anti-FMO5 polyclonal Antibody, 13699-1-AP (Proteintech, Chicago, IL) and then with a horse radish peroxidase-conjugated secondary antibody (Donkey Anti-Rabbit IgG H&L (HRP)), ab97064 (Abcam, Cambridge, MA).

    Techniques: Construct, Clone Assay, Southern Blot, Plasmid Preparation, Knock-Out, Western Blot, Expressing

    Fmo5 −/− mice exhibit a lean phenotype. (A) Body weight plotted as means ± SEM (WT, n = 8; KO, n = 7), * P < 0.05, ** P < 0.01. (B) Internal abdominal view of 30-week-old WT and Fmo5 −/− (KO) mice. (C) Ratio of weight of EWAT to body weight of WT and Fmo5 −/− (KO) mice. 10 week: n = 4 (WT), 3 (KO); 20 week: n = 10 (WT), 5 (KO); 30 week: n = 11 (WT), 8 (KO). Data are expressed as means ± SEM. ** P < 0.01; **** P < 0.0001. (D) Sections of epididymal WAT of 30-week-old WT and Fmo5 −/− (KO) mice stained with haematoxylin and eosin. Scale bar = 100 μm. (E) Food intake expressed as cumulative food intake in grams per g of body weight of WT and Fmo5 −/− (KO) mice. n = 8 (WT), 7 (KO).

    Journal: Biochemical Pharmacology

    Article Title: The phenotype of a knockout mouse identifies flavin-containing monooxygenase 5 (FMO5) as a regulator of metabolic ageing

    doi: 10.1016/j.bcp.2015.05.013

    Figure Lengend Snippet: Fmo5 −/− mice exhibit a lean phenotype. (A) Body weight plotted as means ± SEM (WT, n = 8; KO, n = 7), * P < 0.05, ** P < 0.01. (B) Internal abdominal view of 30-week-old WT and Fmo5 −/− (KO) mice. (C) Ratio of weight of EWAT to body weight of WT and Fmo5 −/− (KO) mice. 10 week: n = 4 (WT), 3 (KO); 20 week: n = 10 (WT), 5 (KO); 30 week: n = 11 (WT), 8 (KO). Data are expressed as means ± SEM. ** P < 0.01; **** P < 0.0001. (D) Sections of epididymal WAT of 30-week-old WT and Fmo5 −/− (KO) mice stained with haematoxylin and eosin. Scale bar = 100 μm. (E) Food intake expressed as cumulative food intake in grams per g of body weight of WT and Fmo5 −/− (KO) mice. n = 8 (WT), 7 (KO).

    Article Snippet: For FMO5, the blot was incubated with Rabbit anti-FMO5 polyclonal Antibody, 13699-1-AP (Proteintech, Chicago, IL) and then with a horse radish peroxidase-conjugated secondary antibody (Donkey Anti-Rabbit IgG H&L (HRP)), ab97064 (Abcam, Cambridge, MA).

    Techniques: Staining

    Fmo5 −/− mice have lower plasma concentrations of glucose, cholesterol and pyruvate. (A) Plasma concentration of glucose in 10- and 30-week-old male and 30-week-old female WT and Fmo5 −/− (KO) mice. 10 week: n = 6 (WT), 3 (KO); 30-week male: n = 19 (WT), 11 (KO); 30-week female: n = 9 (WT), 13 (KO). (B) Plasma concentration of total cholesterol in 10- and 30-week-old male and 30-week-old female WT and Fmo5 −/− (KO) mice. 10 week: n = 11 (WT), 8 (KO); 30-week male: n = 23 (WT), 18 (KO); 30-week female: n = 6 (WT), 10 (KO). (C) Plasma concentration of pyruvate in 15- and 30-week-old male WT and Fmo5 (KO) mice, n = 4 (15 week), 5 (30 week). Data are expressed as means ± SEM. * P < 0.05, ** P < 0.01, **** P < 0.0001.

    Journal: Biochemical Pharmacology

    Article Title: The phenotype of a knockout mouse identifies flavin-containing monooxygenase 5 (FMO5) as a regulator of metabolic ageing

    doi: 10.1016/j.bcp.2015.05.013

    Figure Lengend Snippet: Fmo5 −/− mice have lower plasma concentrations of glucose, cholesterol and pyruvate. (A) Plasma concentration of glucose in 10- and 30-week-old male and 30-week-old female WT and Fmo5 −/− (KO) mice. 10 week: n = 6 (WT), 3 (KO); 30-week male: n = 19 (WT), 11 (KO); 30-week female: n = 9 (WT), 13 (KO). (B) Plasma concentration of total cholesterol in 10- and 30-week-old male and 30-week-old female WT and Fmo5 −/− (KO) mice. 10 week: n = 11 (WT), 8 (KO); 30-week male: n = 23 (WT), 18 (KO); 30-week female: n = 6 (WT), 10 (KO). (C) Plasma concentration of pyruvate in 15- and 30-week-old male WT and Fmo5 (KO) mice, n = 4 (15 week), 5 (30 week). Data are expressed as means ± SEM. * P < 0.05, ** P < 0.01, **** P < 0.0001.

    Article Snippet: For FMO5, the blot was incubated with Rabbit anti-FMO5 polyclonal Antibody, 13699-1-AP (Proteintech, Chicago, IL) and then with a horse radish peroxidase-conjugated secondary antibody (Donkey Anti-Rabbit IgG H&L (HRP)), ab97064 (Abcam, Cambridge, MA).

    Techniques: Concentration Assay

    Fmo5 −/− mice have enhanced energy expenditure, but no increase in voluntary exercise. (A) Energy expenditure in the light, dark and combined light and dark phases. (B) REE. (C) RER for light, dark and combined light and dark phases. Light phase (L, 07:00 to 19:00), dark phase (D, 19:00 to 07:00). Parameters were measured over a 72-h period for 30-week-old male WT ( n = 6) and Fmo5 −/− (KO) ( n = 6) mice. Values are means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. (D) Voluntary wheel running of 30-week-old male WT and Fmo5 −/− (KO) mice was assessed over seven days. Actogram shows average interval count/h ± SEM for a single day.

    Journal: Biochemical Pharmacology

    Article Title: The phenotype of a knockout mouse identifies flavin-containing monooxygenase 5 (FMO5) as a regulator of metabolic ageing

    doi: 10.1016/j.bcp.2015.05.013

    Figure Lengend Snippet: Fmo5 −/− mice have enhanced energy expenditure, but no increase in voluntary exercise. (A) Energy expenditure in the light, dark and combined light and dark phases. (B) REE. (C) RER for light, dark and combined light and dark phases. Light phase (L, 07:00 to 19:00), dark phase (D, 19:00 to 07:00). Parameters were measured over a 72-h period for 30-week-old male WT ( n = 6) and Fmo5 −/− (KO) ( n = 6) mice. Values are means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. (D) Voluntary wheel running of 30-week-old male WT and Fmo5 −/− (KO) mice was assessed over seven days. Actogram shows average interval count/h ± SEM for a single day.

    Article Snippet: For FMO5, the blot was incubated with Rabbit anti-FMO5 polyclonal Antibody, 13699-1-AP (Proteintech, Chicago, IL) and then with a horse radish peroxidase-conjugated secondary antibody (Donkey Anti-Rabbit IgG H&L (HRP)), ab97064 (Abcam, Cambridge, MA).

    Techniques:

    Five key metabolic enzymes are down regulated in the liver of Fmo5 − / − mice. Relative abundance of aldolase B (ALDOB) (A), glycerol 3-phosphate dehydrogenase (GPD1) (B), ketohexokinase (KHK) (C), malic enzyme 1 (ME1) (D) and HMG-CoA synthase 1 (HMGCS1) (E) in liver of WT and Fmo5 −/− (KO) mice determined via proteomic analysis. (F) Western blot analysis of liver lysates of WT and Fmo5 −/− (KO) mice. The blot was incubated with antibodies against ME1 and actin, as a loading control, and developed as described in Section .

    Journal: Biochemical Pharmacology

    Article Title: The phenotype of a knockout mouse identifies flavin-containing monooxygenase 5 (FMO5) as a regulator of metabolic ageing

    doi: 10.1016/j.bcp.2015.05.013

    Figure Lengend Snippet: Five key metabolic enzymes are down regulated in the liver of Fmo5 − / − mice. Relative abundance of aldolase B (ALDOB) (A), glycerol 3-phosphate dehydrogenase (GPD1) (B), ketohexokinase (KHK) (C), malic enzyme 1 (ME1) (D) and HMG-CoA synthase 1 (HMGCS1) (E) in liver of WT and Fmo5 −/− (KO) mice determined via proteomic analysis. (F) Western blot analysis of liver lysates of WT and Fmo5 −/− (KO) mice. The blot was incubated with antibodies against ME1 and actin, as a loading control, and developed as described in Section .

    Article Snippet: For FMO5, the blot was incubated with Rabbit anti-FMO5 polyclonal Antibody, 13699-1-AP (Proteintech, Chicago, IL) and then with a horse radish peroxidase-conjugated secondary antibody (Donkey Anti-Rabbit IgG H&L (HRP)), ab97064 (Abcam, Cambridge, MA).

    Techniques: Western Blot, Incubation

    Relative difference in the abundance of mRNAs in the liver of WT and  Fmo5  − / − mice.

    Journal: Biochemical Pharmacology

    Article Title: The phenotype of a knockout mouse identifies flavin-containing monooxygenase 5 (FMO5) as a regulator of metabolic ageing

    doi: 10.1016/j.bcp.2015.05.013

    Figure Lengend Snippet: Relative difference in the abundance of mRNAs in the liver of WT and Fmo5 − / − mice.

    Article Snippet: For FMO5, the blot was incubated with Rabbit anti-FMO5 polyclonal Antibody, 13699-1-AP (Proteintech, Chicago, IL) and then with a horse radish peroxidase-conjugated secondary antibody (Donkey Anti-Rabbit IgG H&L (HRP)), ab97064 (Abcam, Cambridge, MA).

    Techniques: Expressing, Binding Assay