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mouse anti cdx2  (Biogenex)

 
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    Biogenex mouse anti cdx2
    Mouse Anti Cdx2, supplied by Biogenex, 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/product/cdx2/anti+cdx2+mouse/pm42167122-116-48-50
    Average 86 stars, based on 1 article reviews
    mouse anti cdx2 - by Bioz Stars, 2026-10
    86/100 stars

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    Incubation:

    Article Title: Unraveling the Role of MIXL1 Activation in Endoderm Differentiation of Isogenic Human Induced Pluripotent Stem Cells
    Article Snippet: They were washed with PBS twice and then permeabilised with 0.1% Triton X-100 (Merck) in dPBS (Gibco) (PBST) at RT for 1 hour. .. The organoids were blocked with CAS-Block (Thermo Fisher Scientific) for 90min and then incubated with primary antibody (Sox9 (Merck) 1:500, Ki67 (Abcam) 1:250, CHGA (Novus Biologicals) 1:200, CDX2 (Biogenex) 1:250, Lysozyme (Dako) 1:200) overnight at 4°C. .. Organoids were washed with PBST four times, then incubated with corresponding secondary antibodies and stain (DAPI (1 μg/ml) (Thermo Fisher Scientific) for nuclei in all samples and UEA-1 (Vector Laboratories) 1:200, for select organoids) in CAS-Block at RT for 3 hours.

    Article Title: EPHB2 expression is associated with intestinal phenotype of gastric cancer and indicates better prognosis by suppressing gastric cancer migration
    Article Snippet: Cell lysates were run on a 10% SDS-polyacrylamide gel and transferred to a polyvinylidene fluoride membrane (Millipore Corporation, Bedford, MA, USA). .. The membrane was blocked with 5% non-fat dry milk in PBS-Tween-20 (0.1%, v/v ) for 1 h and then incubated with specific primary antibodies: CDX2 (BioGenex), EPHB2 (R&D systems), HSP90 (Origene), GAPDH (Origene), FAK (Cell signaling, Danvers, MA), and phospho-FAK (Tyr925) (Cell Signaling Technology) overnight at 4°C, washed with TBS containing 0.1% Tween-20, the membrane was incubated for 1 h with secondary antibodies. .. The Alliance-Mini HD9 chemiluminescence documentation system (UVItec Cambridge, UK) was used to visualize the target proteins.

    Article Title: Cdx1 and c-Myc Foster the Initiation of Transdifferentiation of the Normal Esophageal Squamous Epithelium toward Barrett's Esophagus
    Article Snippet: .. Sections were incubated with primary antibody overnight at 4°C and secondary antibody for 30 min at 37°C, and then signal was developed using the DAB substrate kit for peroxidase (Vector Laboratories).Antibodies used were: Cdx1(ab2400, Abcam), Cdx2 (MU392A-UC, Biogenex), Muc2 (VP-M656, Vector Labs), Muc5AC (VP-M657, Vector Labs), Keratin 20 (M7019, DakoCytomation), c-myc (NCL-cMYC, Lab Visions), Keratin 13 (NCL-CK13, Lab Visions), Keratin 19 (TROMA-III, Univ of Iowa Developmental Studies Hybridoma Bank), Keratin 8 (RDI-PRO61038, Research Diagnostics). .. Sections were incubated with primary antibody overnight at 4°C and secondary antibody for 30 min at 37°C, and then signal was developed using the DAB substrate kit for peroxidase (Vector Laboratories).Antibodies used were: Cdx1(ab2400, Abcam), Cdx2 (MU392A-UC, Biogenex), Muc2 (VP-M656, Vector Labs), Muc5AC (VP-M657, Vector Labs), Keratin 20 (M7019, DakoCytomation), c-myc (NCL-cMYC, Lab Visions), Keratin 13 (NCL-CK13, Lab Visions), Keratin 19 (TROMA-III, Univ of Iowa Developmental Studies Hybridoma Bank), Keratin 8 (RDI-PRO61038, Research Diagnostics).

    Article Title: Histone methyltransferases MLL2 and SETD1A/B play distinct roles in H3K4me3 deposition during the transition from totipotency to pluripotency
    Article Snippet: .. Blocking was performed using 5% bovine serum albumin (BSA) in PBS for 2 h before overnight incubation at 4 °C with primary antibodies (H3K4me3 (Abcam, ab8580), MLL2 (Invitrogen, PA5-103371), SETD1A (Abcam, ab70378), SETD1B (Proteintech, 55005-1-AP), KDM5B (Santa, sc-517291), NANOG (Abcam, ab80892), CDX2 (Biogenex, MU392A-5UC) at a 1:200 dilution in 5% BSA. .. Following PVA (0.05% PVA in PBS) washes, samples were exposed to secondary antibodies in 5% BSA for 1 h, employing Dylight 488/549 goat anti-rabbit IgG (Abbkine, A23220 / A23320 ) and Dylight 488 goat anti-mouse IgG (Abbkine, A23210 ; 1:500 dilution) for immunostaining.

    Article Title: Live imaging human embryos reveals mitotic errors and lineage specification prior to implantation
    Article Snippet: Subsequently, embryos were permeabilized with 1X PBS with 0.5% Triton X-100 and then placed in blocking solution (3% BSA (Sigma-Aldrich; 05470-5G) in 1X PBS with 0.2% Triton X-100 (Sigma-Aldrich X100-5ML)) for 2 h at room temperature on a rotating shaker. .. Embryos were then incubated overnight at 4°C on rotating shaker with primary antibodies diluted in blocking solution, at the following concentrations: CDX2 (BioGenex; MU392A-UC) at 1:20, NANOG (2B Scientific, REC-RCAB0001P) at 1:100 and GATA3 (Abcam, ab199428) at dilution 1:100. ..

    Article Title: FOXO1-mediated lipid metabolism maintains mammalian embryos in dormancy.
    Article Snippet: Cells were fixed for 10 min in 4% paraformaldehyde, then permeabilized for 15 min in 0.2% Triton X-100 (Sigma, T8787) in PBS. .. Permeabilized cells were incubated in blocking buffer (0.2% Triton X-100 in PBS, 2% BSA (BSA Fraction V 7.5%, Gibco, 15260-037) and 5% goat serum ( Jackson Immunoresearch/Dianova, 017-000-121)) for 1 h. Cells were incubated overnight at 4 °C with the following primary antibodies: Lamin B1 (Abcam, 16048), H3K9me2 (Abcam, 1220), phospho-mTOR (Abcam, ab131538), pS6 (CST, 4858 S), pAKT (CST,), OCT3/4 (Santa Cruz, sc-5279) and CDX2 (Biogenex, MU392A-UC). .. Cells were washed and incubated with the following secondary antibodies for 1 h at room temperature: donkey anti-rabbit AF647 (Thermo Fisher, A32795) and donkey anti-mouse AF488 (Thermo Fisher, A21202).

    Membrane:

    Article Title: EPHB2 expression is associated with intestinal phenotype of gastric cancer and indicates better prognosis by suppressing gastric cancer migration
    Article Snippet: Cell lysates were run on a 10% SDS-polyacrylamide gel and transferred to a polyvinylidene fluoride membrane (Millipore Corporation, Bedford, MA, USA). .. The membrane was blocked with 5% non-fat dry milk in PBS-Tween-20 (0.1%, v/v ) for 1 h and then incubated with specific primary antibodies: CDX2 (BioGenex), EPHB2 (R&D systems), HSP90 (Origene), GAPDH (Origene), FAK (Cell signaling, Danvers, MA), and phospho-FAK (Tyr925) (Cell Signaling Technology) overnight at 4°C, washed with TBS containing 0.1% Tween-20, the membrane was incubated for 1 h with secondary antibodies. .. The Alliance-Mini HD9 chemiluminescence documentation system (UVItec Cambridge, UK) was used to visualize the target proteins.

    Chromatin Immunoprecipitation:

    Article Title: Elucidating causative gene variants in hereditary Parkinson's disease in the Global Parkinson's Genetics Program (GP2).
    Article Snippet: .. Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used AP2-alpha (Santa Cruz Biotechnology; sc-12726; clone 3B5; 1:200); AP2-gamma (R&D Systems; AF5059; 1:500); AP2-gamma (Santa Cruz Biotechnology, sc-12762; clone 6E4/4; 1:200); Brachyury (R&D Systems; AF2085; 1:500); CDX2 (BioGenex; MU392-UC; clone 4 nature portfolio | reporting sum m ary M arch 2021 CDX2-88; 1:200); CER1 (R&D Systems; AF1075; 1:500); Cytokeratin 7 (Aligent; M7018; clone OV-TL 12/30; 1:100); E-Cadherin (BD Biosciences; 610182; clone 36; 1:200); EOMES (Abcam; ab23345; 1:200); FOXA2 (R&D Systems; AF2400; 1:200); GATA2 (Novus Biologics; NBMP1-82581; 1:200); GATA3 (Abcam; ab199428; clone EPR16651; 1:500); GATA4 (Santa Cruz Biotechnology; sc-25310; clone G-4; 1:200); GATA4 (ThermoFisher Scientific; 14-9980-82; clone eBioEvan; 1:500); GATA6 (R&D Systems; AF1700; 1:500); GATA6 (Cell Signaling Technology; 5851; clone D61E4; 1:2000); GFP (Abcam; ab13970; 1:1000); GFP (Nacalai USA; GF090R; clone GF090R; 1:1000); HAND1 (DSHB; PCRP-HAND1-2A9; clone 2A9 1:200); HNF4-alpha (Abcam; ab201460; clone EPR16885-99; 1:2000); ISL1 (DSHB; PCRP-ISL1-1A9; clone 1A9; 1:100); Laminin (Sigma Aldrich; L9393; 1:200); N-Cadherin (Abcam; ab98952; clone 5D5; 1:200); NANOG (Cell Signaling Technology; 4903; clone D73G4; 1:200); OCT3/4 (Santa Cruz Biotechnology; sc-5279; clone C-10; 1:100); OTX2 (R&D Systems; AF1979; 1:1000); phospho-SMAD1/5 (Cell Signaling Technology; 9516S; clone 41D10; 1:200); Smad2/3 (Cell Signaling Technology; 8685S; clone D7G7; 1:200); SOX17 (R&D Systems; AF1924; 1:500); SOX2 (ThermoFisher Scientific; 14-9811-82; clone Btjce; 1:500); TBX20 (R&D Systems; MAB8124; clone 668710; 1:100); VTCN1 (Abcam; ab209242; clone EPR20236; 1:200). .. AlexaFluor-405 Donkey Anti-Mouse (ThermoFisher Scientific; A48257; 1:500); AlexaFluor-488 Donkey Anti-Rat (ThermoFisher Scientific; A-21208; 1:500); AlexFluor-488 Donkey Anti-Mouse (ThermoFisher Scientific; A-21202; 1:500); AlexaFluor-488 Donkey Anti-Goat (ThermoFisher Scientific; A-11055; 1:500); AlexaFluor-568 Donkey Anti-Rabbit (ThermoFisher Scientific; A10042; 1:500); AlexaFluor-568 Donkey Anti-Rat (ThermoFisher Scientific; A78946; 1:500); AlexaFluor-568 Donkey Anti-Mouse (ThermoFisher Scientific; A10037; 1:500); AlexaFluor-647 Donkey Anti-Rat (ThermoFisher Scientific; A78947; 1:500); AlexaFluor-647 Donkey AntiGoat (ThermoFisher Scientific; A-21447; 1:500); AlexaFluor-647 Donkey Anti-Rabbit (Thermofisher Scientific; A-31573; 1:500); AlexaFluor-647 Donkey Anti-Mouse (ThermoFisher Scientific; A32787; 1:500).

    Flow Cytometry:

    Article Title: Elucidating causative gene variants in hereditary Parkinson's disease in the Global Parkinson's Genetics Program (GP2).
    Article Snippet: .. Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used AP2-alpha (Santa Cruz Biotechnology; sc-12726; clone 3B5; 1:200); AP2-gamma (R&D Systems; AF5059; 1:500); AP2-gamma (Santa Cruz Biotechnology, sc-12762; clone 6E4/4; 1:200); Brachyury (R&D Systems; AF2085; 1:500); CDX2 (BioGenex; MU392-UC; clone 4 nature portfolio | reporting sum m ary M arch 2021 CDX2-88; 1:200); CER1 (R&D Systems; AF1075; 1:500); Cytokeratin 7 (Aligent; M7018; clone OV-TL 12/30; 1:100); E-Cadherin (BD Biosciences; 610182; clone 36; 1:200); EOMES (Abcam; ab23345; 1:200); FOXA2 (R&D Systems; AF2400; 1:200); GATA2 (Novus Biologics; NBMP1-82581; 1:200); GATA3 (Abcam; ab199428; clone EPR16651; 1:500); GATA4 (Santa Cruz Biotechnology; sc-25310; clone G-4; 1:200); GATA4 (ThermoFisher Scientific; 14-9980-82; clone eBioEvan; 1:500); GATA6 (R&D Systems; AF1700; 1:500); GATA6 (Cell Signaling Technology; 5851; clone D61E4; 1:2000); GFP (Abcam; ab13970; 1:1000); GFP (Nacalai USA; GF090R; clone GF090R; 1:1000); HAND1 (DSHB; PCRP-HAND1-2A9; clone 2A9 1:200); HNF4-alpha (Abcam; ab201460; clone EPR16885-99; 1:2000); ISL1 (DSHB; PCRP-ISL1-1A9; clone 1A9; 1:100); Laminin (Sigma Aldrich; L9393; 1:200); N-Cadherin (Abcam; ab98952; clone 5D5; 1:200); NANOG (Cell Signaling Technology; 4903; clone D73G4; 1:200); OCT3/4 (Santa Cruz Biotechnology; sc-5279; clone C-10; 1:100); OTX2 (R&D Systems; AF1979; 1:1000); phospho-SMAD1/5 (Cell Signaling Technology; 9516S; clone 41D10; 1:200); Smad2/3 (Cell Signaling Technology; 8685S; clone D7G7; 1:200); SOX17 (R&D Systems; AF1924; 1:500); SOX2 (ThermoFisher Scientific; 14-9811-82; clone Btjce; 1:500); TBX20 (R&D Systems; MAB8124; clone 668710; 1:100); VTCN1 (Abcam; ab209242; clone EPR20236; 1:200). .. AlexaFluor-405 Donkey Anti-Mouse (ThermoFisher Scientific; A48257; 1:500); AlexaFluor-488 Donkey Anti-Rat (ThermoFisher Scientific; A-21208; 1:500); AlexFluor-488 Donkey Anti-Mouse (ThermoFisher Scientific; A-21202; 1:500); AlexaFluor-488 Donkey Anti-Goat (ThermoFisher Scientific; A-11055; 1:500); AlexaFluor-568 Donkey Anti-Rabbit (ThermoFisher Scientific; A10042; 1:500); AlexaFluor-568 Donkey Anti-Rat (ThermoFisher Scientific; A78946; 1:500); AlexaFluor-568 Donkey Anti-Mouse (ThermoFisher Scientific; A10037; 1:500); AlexaFluor-647 Donkey Anti-Rat (ThermoFisher Scientific; A78947; 1:500); AlexaFluor-647 Donkey AntiGoat (ThermoFisher Scientific; A-21447; 1:500); AlexaFluor-647 Donkey Anti-Rabbit (Thermofisher Scientific; A-31573; 1:500); AlexaFluor-647 Donkey Anti-Mouse (ThermoFisher Scientific; A32787; 1:500).

    Magnetic Resonance Imaging:

    Article Title: Elucidating causative gene variants in hereditary Parkinson's disease in the Global Parkinson's Genetics Program (GP2).
    Article Snippet: .. Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used AP2-alpha (Santa Cruz Biotechnology; sc-12726; clone 3B5; 1:200); AP2-gamma (R&D Systems; AF5059; 1:500); AP2-gamma (Santa Cruz Biotechnology, sc-12762; clone 6E4/4; 1:200); Brachyury (R&D Systems; AF2085; 1:500); CDX2 (BioGenex; MU392-UC; clone 4 nature portfolio | reporting sum m ary M arch 2021 CDX2-88; 1:200); CER1 (R&D Systems; AF1075; 1:500); Cytokeratin 7 (Aligent; M7018; clone OV-TL 12/30; 1:100); E-Cadherin (BD Biosciences; 610182; clone 36; 1:200); EOMES (Abcam; ab23345; 1:200); FOXA2 (R&D Systems; AF2400; 1:200); GATA2 (Novus Biologics; NBMP1-82581; 1:200); GATA3 (Abcam; ab199428; clone EPR16651; 1:500); GATA4 (Santa Cruz Biotechnology; sc-25310; clone G-4; 1:200); GATA4 (ThermoFisher Scientific; 14-9980-82; clone eBioEvan; 1:500); GATA6 (R&D Systems; AF1700; 1:500); GATA6 (Cell Signaling Technology; 5851; clone D61E4; 1:2000); GFP (Abcam; ab13970; 1:1000); GFP (Nacalai USA; GF090R; clone GF090R; 1:1000); HAND1 (DSHB; PCRP-HAND1-2A9; clone 2A9 1:200); HNF4-alpha (Abcam; ab201460; clone EPR16885-99; 1:2000); ISL1 (DSHB; PCRP-ISL1-1A9; clone 1A9; 1:100); Laminin (Sigma Aldrich; L9393; 1:200); N-Cadherin (Abcam; ab98952; clone 5D5; 1:200); NANOG (Cell Signaling Technology; 4903; clone D73G4; 1:200); OCT3/4 (Santa Cruz Biotechnology; sc-5279; clone C-10; 1:100); OTX2 (R&D Systems; AF1979; 1:1000); phospho-SMAD1/5 (Cell Signaling Technology; 9516S; clone 41D10; 1:200); Smad2/3 (Cell Signaling Technology; 8685S; clone D7G7; 1:200); SOX17 (R&D Systems; AF1924; 1:500); SOX2 (ThermoFisher Scientific; 14-9811-82; clone Btjce; 1:500); TBX20 (R&D Systems; MAB8124; clone 668710; 1:100); VTCN1 (Abcam; ab209242; clone EPR20236; 1:200). .. AlexaFluor-405 Donkey Anti-Mouse (ThermoFisher Scientific; A48257; 1:500); AlexaFluor-488 Donkey Anti-Rat (ThermoFisher Scientific; A-21208; 1:500); AlexFluor-488 Donkey Anti-Mouse (ThermoFisher Scientific; A-21202; 1:500); AlexaFluor-488 Donkey Anti-Goat (ThermoFisher Scientific; A-11055; 1:500); AlexaFluor-568 Donkey Anti-Rabbit (ThermoFisher Scientific; A10042; 1:500); AlexaFluor-568 Donkey Anti-Rat (ThermoFisher Scientific; A78946; 1:500); AlexaFluor-568 Donkey Anti-Mouse (ThermoFisher Scientific; A10037; 1:500); AlexaFluor-647 Donkey Anti-Rat (ThermoFisher Scientific; A78947; 1:500); AlexaFluor-647 Donkey AntiGoat (ThermoFisher Scientific; A-21447; 1:500); AlexaFluor-647 Donkey Anti-Rabbit (Thermofisher Scientific; A-31573; 1:500); AlexaFluor-647 Donkey Anti-Mouse (ThermoFisher Scientific; A32787; 1:500).

    other:

    Article Title: Embryo-uterine interaction coordinates mouse embryogenesis during implantation.
    Article Snippet: Embryo implantation into the uterus marks a key transition in mammalian development.. In mice, implantation is mediated by the trophoblast and is accompanied by a morphological transition from the blastocyst to the egg cylinder.. However, the roles of trophoblast-uterine interactions in embryo morphogenesis during implantation are poorly understood due to inaccessibility in utero and the remaining challenges to recapitulate it ex vivo from the blastocyst.

    Blocking Assay:

    Article Title: Histone methyltransferases MLL2 and SETD1A/B play distinct roles in H3K4me3 deposition during the transition from totipotency to pluripotency
    Article Snippet: .. Blocking was performed using 5% bovine serum albumin (BSA) in PBS for 2 h before overnight incubation at 4 °C with primary antibodies (H3K4me3 (Abcam, ab8580), MLL2 (Invitrogen, PA5-103371), SETD1A (Abcam, ab70378), SETD1B (Proteintech, 55005-1-AP), KDM5B (Santa, sc-517291), NANOG (Abcam, ab80892), CDX2 (Biogenex, MU392A-5UC) at a 1:200 dilution in 5% BSA. .. Following PVA (0.05% PVA in PBS) washes, samples were exposed to secondary antibodies in 5% BSA for 1 h, employing Dylight 488/549 goat anti-rabbit IgG (Abbkine, A23220 / A23320 ) and Dylight 488 goat anti-mouse IgG (Abbkine, A23210 ; 1:500 dilution) for immunostaining.

    Article Title: Live imaging human embryos reveals mitotic errors and lineage specification prior to implantation
    Article Snippet: Subsequently, embryos were permeabilized with 1X PBS with 0.5% Triton X-100 and then placed in blocking solution (3% BSA (Sigma-Aldrich; 05470-5G) in 1X PBS with 0.2% Triton X-100 (Sigma-Aldrich X100-5ML)) for 2 h at room temperature on a rotating shaker. .. Embryos were then incubated overnight at 4°C on rotating shaker with primary antibodies diluted in blocking solution, at the following concentrations: CDX2 (BioGenex; MU392A-UC) at 1:20, NANOG (2B Scientific, REC-RCAB0001P) at 1:100 and GATA3 (Abcam, ab199428) at dilution 1:100. ..

    Article Title: FOXO1-mediated lipid metabolism maintains mammalian embryos in dormancy.
    Article Snippet: Cells were fixed for 10 min in 4% paraformaldehyde, then permeabilized for 15 min in 0.2% Triton X-100 (Sigma, T8787) in PBS. .. Permeabilized cells were incubated in blocking buffer (0.2% Triton X-100 in PBS, 2% BSA (BSA Fraction V 7.5%, Gibco, 15260-037) and 5% goat serum ( Jackson Immunoresearch/Dianova, 017-000-121)) for 1 h. Cells were incubated overnight at 4 °C with the following primary antibodies: Lamin B1 (Abcam, 16048), H3K9me2 (Abcam, 1220), phospho-mTOR (Abcam, ab131538), pS6 (CST, 4858 S), pAKT (CST,), OCT3/4 (Santa Cruz, sc-5279) and CDX2 (Biogenex, MU392A-UC). .. Cells were washed and incubated with the following secondary antibodies for 1 h at room temperature: donkey anti-rabbit AF647 (Thermo Fisher, A32795) and donkey anti-mouse AF488 (Thermo Fisher, A21202).



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    (A) Breeding strategy for the production of CEC- Acly −/− / Acss 2 −/− double knockout (DKO) mice; (B) Body weight changes in Acly f/f / Acss2 f/f <t>;Cdx2-CreERT2</t> mice following tamoxifen (TAM) treatment with or without acetate (Ac) supplementation; (C) Kaplan-Meier survival curves of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) treatment with or without acetate supplementation; (D) Western blot assessment of senescence hallmarks in colonic crypt cells from Acly f/f / Acss2 f/f (Ctrl) and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 (DKO) mice on day 13 following TAM and acetate treatment; (E) RT-qPCR quantitation of SASP cytokines in colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (F) Serum cytokine concentrations in Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (G) H&E stained lung sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM treatment; (H) Quantitation of fecal SCFAs (acetate, propionate, butyrate) in Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (I) Alcian blue staining of colon sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (J) Confocal microscopic images of ψH2AX immunofluorescence staining in colonic sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment. Arrows indicate ψH2AX loci in the nucleus of crypt epithelial cells; (K) Quantitation of ψH2AX-positive cells in colonic mucosa of Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (L) Quantitation of acetyl-CoA concentrations in total cell lysates and nuclear, cytosolic and mitochondrial fractions of colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment. Data are presented as mean ± SD. *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001 by two-way ANOVA.
    Cdx2 Creert2 Transgenic Mice, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cdx2  (Cell Marque)
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    Cell Marque cdx2
    (A) Breeding strategy for the production of CEC- Acly −/− / Acss 2 −/− double knockout (DKO) mice; (B) Body weight changes in Acly f/f / Acss2 f/f <t>;Cdx2-CreERT2</t> mice following tamoxifen (TAM) treatment with or without acetate (Ac) supplementation; (C) Kaplan-Meier survival curves of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) treatment with or without acetate supplementation; (D) Western blot assessment of senescence hallmarks in colonic crypt cells from Acly f/f / Acss2 f/f (Ctrl) and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 (DKO) mice on day 13 following TAM and acetate treatment; (E) RT-qPCR quantitation of SASP cytokines in colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (F) Serum cytokine concentrations in Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (G) H&E stained lung sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM treatment; (H) Quantitation of fecal SCFAs (acetate, propionate, butyrate) in Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (I) Alcian blue staining of colon sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (J) Confocal microscopic images of ψH2AX immunofluorescence staining in colonic sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment. Arrows indicate ψH2AX loci in the nucleus of crypt epithelial cells; (K) Quantitation of ψH2AX-positive cells in colonic mucosa of Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (L) Quantitation of acetyl-CoA concentrations in total cell lysates and nuclear, cytosolic and mitochondrial fractions of colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment. Data are presented as mean ± SD. *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001 by two-way ANOVA.
    Cdx2, supplied by Cell Marque, 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/product/cdx2/cdx2+colon+markers/pm42120420-272-3-4
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    (A) Breeding strategy for the production of CEC- Acly −/− / Acss 2 −/− double knockout (DKO) mice; (B) Body weight changes in Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) treatment with or without acetate (Ac) supplementation; (C) Kaplan-Meier survival curves of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) treatment with or without acetate supplementation; (D) Western blot assessment of senescence hallmarks in colonic crypt cells from Acly f/f / Acss2 f/f (Ctrl) and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 (DKO) mice on day 13 following TAM and acetate treatment; (E) RT-qPCR quantitation of SASP cytokines in colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (F) Serum cytokine concentrations in Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (G) H&E stained lung sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM treatment; (H) Quantitation of fecal SCFAs (acetate, propionate, butyrate) in Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (I) Alcian blue staining of colon sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (J) Confocal microscopic images of ψH2AX immunofluorescence staining in colonic sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment. Arrows indicate ψH2AX loci in the nucleus of crypt epithelial cells; (K) Quantitation of ψH2AX-positive cells in colonic mucosa of Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (L) Quantitation of acetyl-CoA concentrations in total cell lysates and nuclear, cytosolic and mitochondrial fractions of colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment. Data are presented as mean ± SD. *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001 by two-way ANOVA.

    Journal: bioRxiv

    Article Title: Gut Microbiota-derived Acetate Safeguards the Colonic Epithelial Acetyl-CoA Reserve to Avert Colonic Senescence

    doi: 10.64898/2026.05.15.725523

    Figure Lengend Snippet: (A) Breeding strategy for the production of CEC- Acly −/− / Acss 2 −/− double knockout (DKO) mice; (B) Body weight changes in Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) treatment with or without acetate (Ac) supplementation; (C) Kaplan-Meier survival curves of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) treatment with or without acetate supplementation; (D) Western blot assessment of senescence hallmarks in colonic crypt cells from Acly f/f / Acss2 f/f (Ctrl) and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 (DKO) mice on day 13 following TAM and acetate treatment; (E) RT-qPCR quantitation of SASP cytokines in colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (F) Serum cytokine concentrations in Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (G) H&E stained lung sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM treatment; (H) Quantitation of fecal SCFAs (acetate, propionate, butyrate) in Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (I) Alcian blue staining of colon sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (J) Confocal microscopic images of ψH2AX immunofluorescence staining in colonic sections from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment. Arrows indicate ψH2AX loci in the nucleus of crypt epithelial cells; (K) Quantitation of ψH2AX-positive cells in colonic mucosa of Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment; (L) Quantitation of acetyl-CoA concentrations in total cell lysates and nuclear, cytosolic and mitochondrial fractions of colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and acetate treatment. Data are presented as mean ± SD. *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001 by two-way ANOVA.

    Article Snippet: CDX2-Cre transgenic mice (B6.Cg-Tg(CDX2-cre)101Erf/J; Stock # 009350), CDX2-CreERT2 transgenic mice (B6.Cg-Tg(CDX2-Cre/ERT2)752Erf/J; Stock # 022390), p16-3MR transgenic mice (B6.Cg-Tg(Cdkn2a/luc/RFP/TK)1Cmps/J, Stock # 037045) and Trp53 −/− mice (B6.129S2- Trp53 tm1Tyj /J; Stock # 002101) were purchased from Jackson Laboratory.

    Techniques: Double Knockout, Western Blot, Quantitative RT-PCR, Quantitation Assay, Staining, Immunofluorescence

    (A) Schematic illustration for the derivation of Acly -deleted colonic organoids from Acly f/f ;Cdx2-CreERT2 mice; (B) SA-β-gal expression in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-hydroxytamoxyfen (4-OHT) treatment; (C) Western blot assessment of canonical senescence hallmarks in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-OHT treatment; (D) RT-qPCR quantitation of SASP cytokine expression in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-OHT treatment; *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001 by two-way ANOVA. (E) Western blot analysis of IL-6 and TNF-α expression in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-OHT treatment; (F) Western blot analysis of senescence-related biomarkers in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-OHT treatment; (G) Western blot analysis of senescence hallmarks in Acly f/f ;Cdx2-CreERT2 colonic organoids treated with 4-OHT with or without acetate (Ac) supplementation in the culture media; (H) Human colonic organoids transduced with lentivirus carrying control shRNA (shCtrl) or ACLY-specific shRNA (shACLY) and cultured in the presence or absence of acetate (Ac) supplementation. Note the unique senescent morphology of shACLY-transduced organoids; (I) Western blot confirmation of ACLY knockdown, induction of senescence hallmarks, and suppression of these hallmarks by Ac supplementation in human organoids transduced with shACLY-lentivirus.

    Journal: bioRxiv

    Article Title: Gut Microbiota-derived Acetate Safeguards the Colonic Epithelial Acetyl-CoA Reserve to Avert Colonic Senescence

    doi: 10.64898/2026.05.15.725523

    Figure Lengend Snippet: (A) Schematic illustration for the derivation of Acly -deleted colonic organoids from Acly f/f ;Cdx2-CreERT2 mice; (B) SA-β-gal expression in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-hydroxytamoxyfen (4-OHT) treatment; (C) Western blot assessment of canonical senescence hallmarks in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-OHT treatment; (D) RT-qPCR quantitation of SASP cytokine expression in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-OHT treatment; *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001 by two-way ANOVA. (E) Western blot analysis of IL-6 and TNF-α expression in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-OHT treatment; (F) Western blot analysis of senescence-related biomarkers in Acly f/f ;Cdx2-CreERT2 colonic organoids on days 0, 4 and 6 after 4-OHT treatment; (G) Western blot analysis of senescence hallmarks in Acly f/f ;Cdx2-CreERT2 colonic organoids treated with 4-OHT with or without acetate (Ac) supplementation in the culture media; (H) Human colonic organoids transduced with lentivirus carrying control shRNA (shCtrl) or ACLY-specific shRNA (shACLY) and cultured in the presence or absence of acetate (Ac) supplementation. Note the unique senescent morphology of shACLY-transduced organoids; (I) Western blot confirmation of ACLY knockdown, induction of senescence hallmarks, and suppression of these hallmarks by Ac supplementation in human organoids transduced with shACLY-lentivirus.

    Article Snippet: CDX2-Cre transgenic mice (B6.Cg-Tg(CDX2-cre)101Erf/J; Stock # 009350), CDX2-CreERT2 transgenic mice (B6.Cg-Tg(CDX2-Cre/ERT2)752Erf/J; Stock # 022390), p16-3MR transgenic mice (B6.Cg-Tg(Cdkn2a/luc/RFP/TK)1Cmps/J, Stock # 037045) and Trp53 −/− mice (B6.129S2- Trp53 tm1Tyj /J; Stock # 002101) were purchased from Jackson Laboratory.

    Techniques: Expressing, Western Blot, Quantitative RT-PCR, Quantitation Assay, Transduction, Control, shRNA, Cell Culture, Knockdown

    (A) Acetyl-protein profiles revealed by immunoprecipitation with anti-acetyl-lysine antibody in purified colonic crypt cells from Acly f/f and CEC- Acly −/− mice on day 7 following ABX and acetate (Ac) feeding; (B) Kaplan-Meier survival curves of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) and SAHA treatment; (C) Acetyl-protein profiles in colonic crypt cells of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and SAHA treatment; (D) Western blot assessment of senescence hallmarks in purified colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and SAHA treatment; (E) Schematic illustration of acetyl-proteomics procedure; (F) Heatmap showing changes in specific lysine acetylation in a cohort of nuclear and mitochondrial proteins identified by acetyl-proteomics in CEC- Acly −/− colonic crypt cells with or without ABX; (G) Western blot analysis of H4 acetylation at K8 and K16 in purified colonic crypt cells from Acly f/f and CEC- Acly −/− mice on day 7 following ABX and Ac feeding; (H) Immunoprecipitation analysis of nuclear protein acetylation using anti-acetyl-lysine antibody in purified colonic crypt cells from Acly f/f and CEC- Acly −/− mice on day 7 following ABX and Ac feeding; (I) Immunoprecipitation analysis of mitochondrial protein acetylation using anti-acetyl-lysine antibody in purified colonic crypt cells from Acly f/f and CEC- Acly −/− mice on day 7 following ABX and Ac feeding.

    Journal: bioRxiv

    Article Title: Gut Microbiota-derived Acetate Safeguards the Colonic Epithelial Acetyl-CoA Reserve to Avert Colonic Senescence

    doi: 10.64898/2026.05.15.725523

    Figure Lengend Snippet: (A) Acetyl-protein profiles revealed by immunoprecipitation with anti-acetyl-lysine antibody in purified colonic crypt cells from Acly f/f and CEC- Acly −/− mice on day 7 following ABX and acetate (Ac) feeding; (B) Kaplan-Meier survival curves of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) and SAHA treatment; (C) Acetyl-protein profiles in colonic crypt cells of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and SAHA treatment; (D) Western blot assessment of senescence hallmarks in purified colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and SAHA treatment; (E) Schematic illustration of acetyl-proteomics procedure; (F) Heatmap showing changes in specific lysine acetylation in a cohort of nuclear and mitochondrial proteins identified by acetyl-proteomics in CEC- Acly −/− colonic crypt cells with or without ABX; (G) Western blot analysis of H4 acetylation at K8 and K16 in purified colonic crypt cells from Acly f/f and CEC- Acly −/− mice on day 7 following ABX and Ac feeding; (H) Immunoprecipitation analysis of nuclear protein acetylation using anti-acetyl-lysine antibody in purified colonic crypt cells from Acly f/f and CEC- Acly −/− mice on day 7 following ABX and Ac feeding; (I) Immunoprecipitation analysis of mitochondrial protein acetylation using anti-acetyl-lysine antibody in purified colonic crypt cells from Acly f/f and CEC- Acly −/− mice on day 7 following ABX and Ac feeding.

    Article Snippet: CDX2-Cre transgenic mice (B6.Cg-Tg(CDX2-cre)101Erf/J; Stock # 009350), CDX2-CreERT2 transgenic mice (B6.Cg-Tg(CDX2-Cre/ERT2)752Erf/J; Stock # 022390), p16-3MR transgenic mice (B6.Cg-Tg(Cdkn2a/luc/RFP/TK)1Cmps/J, Stock # 037045) and Trp53 −/− mice (B6.129S2- Trp53 tm1Tyj /J; Stock # 002101) were purchased from Jackson Laboratory.

    Techniques: Immunoprecipitation, Purification, Western Blot

    (A) Acetyl-protein profiles in Acly f/f ;Cdx2-CreERT2 colonic organoids on day 6 after 4-OHT treatment with or without acetate supplementation in the media; (B) Western blot analysis of senescence markers in Acly f/f ;Cdx2-CreERT2 colonic organoids treated with 4-OHT and Vorinostat (SAHA); (C) Western blot analysis of H4 acetylation in colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice treated with TAM and SAHA; (D) Immunoprecipitation analysis of nuclear protein acetylation using anti-acetyl-lysine antibody in colonic crypt cells prepared from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice treated with TAM and SAHA; (E) Immunoprecipitation analysis of mitochondrial protein acetylation using anti-acetyl-lysine antibody in colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice treated with TAM and SAHA; (F) FACS analysis of JC-1 stained Acly f/f ;Cdx2-CreERT2 colonic organoids on day 6 after 4-OHT treatment; (G) Mitochondrial membrane potentials in Acly f/f ;Cdx2-CreERT2 colonic organoids on day 6 after 4-OHT treatment, expressed as red (Q2) to green (Q3) ratio based on the FACS data; (H) Cellular ATP concentration in purified colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 after TAM and acetate treatment; (I) Cellular ROS production in purified colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 after TAM and acetate treatment; (J) Western blot assessment of senescence markers in Acly f/f ;Cdx2-CreERT2 colonic organoids on day 6 after 4-OHT and NAC treatment. (K) Molecular mechanism whereby acetyl-CoA deficiency triggers colonic senescence. Colonic epithelial Acly ablation combined with intestinal bacterial depletion or Acss2 ablation causes epithelial acetyl-CoA deficiency, which depletes lysine acetylation in a cohort of nuclear and mitochondrial proteins. Particularly, the depletion of ATP5F1A acetylation at K161 disrupts ATP synthesis and the depletion of H4 acetylation at K5 and K8 disrupts its DNA repair activity. These lead to increased oxidative stress and increased DNA damage response, which together trigger robust colonic epithelial senescence. Widespread colonic senescence causes severe systemic inflammation and premature death of the mice.

    Journal: bioRxiv

    Article Title: Gut Microbiota-derived Acetate Safeguards the Colonic Epithelial Acetyl-CoA Reserve to Avert Colonic Senescence

    doi: 10.64898/2026.05.15.725523

    Figure Lengend Snippet: (A) Acetyl-protein profiles in Acly f/f ;Cdx2-CreERT2 colonic organoids on day 6 after 4-OHT treatment with or without acetate supplementation in the media; (B) Western blot analysis of senescence markers in Acly f/f ;Cdx2-CreERT2 colonic organoids treated with 4-OHT and Vorinostat (SAHA); (C) Western blot analysis of H4 acetylation in colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice treated with TAM and SAHA; (D) Immunoprecipitation analysis of nuclear protein acetylation using anti-acetyl-lysine antibody in colonic crypt cells prepared from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice treated with TAM and SAHA; (E) Immunoprecipitation analysis of mitochondrial protein acetylation using anti-acetyl-lysine antibody in colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice treated with TAM and SAHA; (F) FACS analysis of JC-1 stained Acly f/f ;Cdx2-CreERT2 colonic organoids on day 6 after 4-OHT treatment; (G) Mitochondrial membrane potentials in Acly f/f ;Cdx2-CreERT2 colonic organoids on day 6 after 4-OHT treatment, expressed as red (Q2) to green (Q3) ratio based on the FACS data; (H) Cellular ATP concentration in purified colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 after TAM and acetate treatment; (I) Cellular ROS production in purified colonic crypt cells from Acly f/f / Acss2 f/f and Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 after TAM and acetate treatment; (J) Western blot assessment of senescence markers in Acly f/f ;Cdx2-CreERT2 colonic organoids on day 6 after 4-OHT and NAC treatment. (K) Molecular mechanism whereby acetyl-CoA deficiency triggers colonic senescence. Colonic epithelial Acly ablation combined with intestinal bacterial depletion or Acss2 ablation causes epithelial acetyl-CoA deficiency, which depletes lysine acetylation in a cohort of nuclear and mitochondrial proteins. Particularly, the depletion of ATP5F1A acetylation at K161 disrupts ATP synthesis and the depletion of H4 acetylation at K5 and K8 disrupts its DNA repair activity. These lead to increased oxidative stress and increased DNA damage response, which together trigger robust colonic epithelial senescence. Widespread colonic senescence causes severe systemic inflammation and premature death of the mice.

    Article Snippet: CDX2-Cre transgenic mice (B6.Cg-Tg(CDX2-cre)101Erf/J; Stock # 009350), CDX2-CreERT2 transgenic mice (B6.Cg-Tg(CDX2-Cre/ERT2)752Erf/J; Stock # 022390), p16-3MR transgenic mice (B6.Cg-Tg(Cdkn2a/luc/RFP/TK)1Cmps/J, Stock # 037045) and Trp53 −/− mice (B6.129S2- Trp53 tm1Tyj /J; Stock # 002101) were purchased from Jackson Laboratory.

    Techniques: Western Blot, Immunoprecipitation, Staining, Membrane, Concentration Assay, Purification, Activity Assay

    (A) Schematic illustration of an organoid rescue strategy using recombinant lentivirus; (B) Microscopic images of Acly f/f ;Cdx2-CreERT2 colonic organoids transduced with recombinant GFP-lentivirus carrying cDNA as indicated. Unless indicated, all organoids were treated with 4-OHT to deplete Acly . GFP fluorescence confirms the success in organoid transduction, and the normal and senescent morphology of the same organoids are identified by regular light microscopy; V, vector. (C,D) Western blot assessment of the induction or suppression of senescence hallmarks in organoids transduced with a recombinant lentivirus indicated on the top. Overexpression of each corresponding protein confirms the successful transduction of the organoids; EV: Empty vector. (E) Schematic illustration of acetylation of the lysine (K) residues identified by acetyl-proteomics in H4 and ATP5F1A, their mutation to glutamine (Q) or arginine (R) in each construct, and the outcome of the rescue experiments; (F) Microscopic images of Acly f/f ;Cdx2-CreERT2 colonic organoids transduced with a recombinant GFP-lentivirus carrying wildtype H4c1 or Atp5f1a cDNA, or their mutants in which all four K residues were converted to Q or R. (G) Microscopic images of Acly f/f ;Cdx2-CreERT2 colonic organoids transduced with a recombinant GFP-lentivirus carrying one of the H4c1 double mutants in which two K residues were mutated to R, or carrying one of the Atp5f1a single mutants in which each K residue was individually converted to R; (H) Western blot assessment of senescence hallmarks in Acly f/f ;Cdx2-CreERT2 organoids transduced with a recombinant lentivirus carrying wildtype H4c1, H4c1(4K>4Q) or H4c1(4K>4R) mutant, or carrying wildtype Atp5f1a, Atp5f1a(4K>4Q) or Atp5f1a(4K>5R) mutant; (I) Western blot assessment of senescence hallmarks in Acly f/f ;Cdx2-CreERT2 organoids transduced with a recombinant lentivirus carrying one of the H4c1 double mutants or one of the Atp5f1a single mutants as indicated; (J) Quantitation of cellular ATP concentration in Acly f/f ;Cdx2-CreERT2 organoids transduced with indicated recombinant GFP-lentivirus constructs; (I) Quantitation of cellular ROS production in Acly f/f ;Cdx2-CreERT2 organoids transduced with indicated recombinant GFP-lentivirus constructs; (L) Kaplan-Meier survival curves of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) and N-acetylcysteine (NAC) treatment; (M) Quantitation of ROS production in purified colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and NAC treatment; (N) Western blot assessment of senescence hallmarks in purified colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and NAC treatment; (O) RT-qPCR quantitation of SASP cytokines in purified colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and NAC treatment.

    Journal: bioRxiv

    Article Title: Gut Microbiota-derived Acetate Safeguards the Colonic Epithelial Acetyl-CoA Reserve to Avert Colonic Senescence

    doi: 10.64898/2026.05.15.725523

    Figure Lengend Snippet: (A) Schematic illustration of an organoid rescue strategy using recombinant lentivirus; (B) Microscopic images of Acly f/f ;Cdx2-CreERT2 colonic organoids transduced with recombinant GFP-lentivirus carrying cDNA as indicated. Unless indicated, all organoids were treated with 4-OHT to deplete Acly . GFP fluorescence confirms the success in organoid transduction, and the normal and senescent morphology of the same organoids are identified by regular light microscopy; V, vector. (C,D) Western blot assessment of the induction or suppression of senescence hallmarks in organoids transduced with a recombinant lentivirus indicated on the top. Overexpression of each corresponding protein confirms the successful transduction of the organoids; EV: Empty vector. (E) Schematic illustration of acetylation of the lysine (K) residues identified by acetyl-proteomics in H4 and ATP5F1A, their mutation to glutamine (Q) or arginine (R) in each construct, and the outcome of the rescue experiments; (F) Microscopic images of Acly f/f ;Cdx2-CreERT2 colonic organoids transduced with a recombinant GFP-lentivirus carrying wildtype H4c1 or Atp5f1a cDNA, or their mutants in which all four K residues were converted to Q or R. (G) Microscopic images of Acly f/f ;Cdx2-CreERT2 colonic organoids transduced with a recombinant GFP-lentivirus carrying one of the H4c1 double mutants in which two K residues were mutated to R, or carrying one of the Atp5f1a single mutants in which each K residue was individually converted to R; (H) Western blot assessment of senescence hallmarks in Acly f/f ;Cdx2-CreERT2 organoids transduced with a recombinant lentivirus carrying wildtype H4c1, H4c1(4K>4Q) or H4c1(4K>4R) mutant, or carrying wildtype Atp5f1a, Atp5f1a(4K>4Q) or Atp5f1a(4K>5R) mutant; (I) Western blot assessment of senescence hallmarks in Acly f/f ;Cdx2-CreERT2 organoids transduced with a recombinant lentivirus carrying one of the H4c1 double mutants or one of the Atp5f1a single mutants as indicated; (J) Quantitation of cellular ATP concentration in Acly f/f ;Cdx2-CreERT2 organoids transduced with indicated recombinant GFP-lentivirus constructs; (I) Quantitation of cellular ROS production in Acly f/f ;Cdx2-CreERT2 organoids transduced with indicated recombinant GFP-lentivirus constructs; (L) Kaplan-Meier survival curves of Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice following tamoxifen (TAM) and N-acetylcysteine (NAC) treatment; (M) Quantitation of ROS production in purified colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and NAC treatment; (N) Western blot assessment of senescence hallmarks in purified colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and NAC treatment; (O) RT-qPCR quantitation of SASP cytokines in purified colonic crypt cells from Acly f/f / Acss2 f/f ;Cdx2-CreERT2 mice on day 13 following TAM and NAC treatment.

    Article Snippet: CDX2-Cre transgenic mice (B6.Cg-Tg(CDX2-cre)101Erf/J; Stock # 009350), CDX2-CreERT2 transgenic mice (B6.Cg-Tg(CDX2-Cre/ERT2)752Erf/J; Stock # 022390), p16-3MR transgenic mice (B6.Cg-Tg(Cdkn2a/luc/RFP/TK)1Cmps/J, Stock # 037045) and Trp53 −/− mice (B6.129S2- Trp53 tm1Tyj /J; Stock # 002101) were purchased from Jackson Laboratory.

    Techniques: Recombinant, Transduction, Fluorescence, Light Microscopy, Plasmid Preparation, Western Blot, Over Expression, Mutagenesis, Construct, Residue, Quantitation Assay, Concentration Assay, Purification, Quantitative RT-PCR