Review



p53 luciferase reporter construct  (Addgene inc)


Bioz Verified Symbol Addgene inc is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 92
      Buy from Supplier

    Structured Review

    Addgene inc p53 luciferase reporter construct
    P53 Luciferase Reporter Construct, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p53+luciferase+reporter+construct/GFP-p53+(NES-)+(Plasmid+%2312092)/pm38512421-113-34-58
    Average 92 stars, based on 18 article reviews
    p53 luciferase reporter construct - by Bioz Stars, 2026-10
    92/100 stars

    Images

    Related Articles

    Luciferase:

    Article Title: HIPK2 C-terminal domain inhibits NF-κB signaling and renal inflammation in kidney injury.
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, #SL100688). .. Luciferase reporter gene assays HEK293T cells were co-transfected with pcDNA4B or HIPK2CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Dr. Huabao Xiong (28), and SBE-luc and p53-luc were obtained from Addgene). ..

    Article Title: HIPK2 C-terminal domain inhibits NF- κ B signaling and renal inflammation in kidney injury
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, SL100688). .. HEK293T cells were cotransfected with pcDNA4B or HIPK2-CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Huabao Xiong [Department of Medicine, Precision Immunology Institute, Icahn School of Medicine at Mount Sinai], ref. , and SBE-luc and p53-luc were obtained from Addgene). ..

    Article Title: Inhibition of Kpn β 1 Mediated Nuclear Import Enhances Cisplatin Chemosensitivity in Cervical Cancer.
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted uorescence microscope under 100 X oil immersion. .. Page 6/23 Luciferase reporter assay SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing ve copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository(26) and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 hours, followed by 30 μM cisplatin for 24 hours, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: Inhibition of Kpnβ1 mediated nuclear import enhances cisplatin chemosensitivity in cervical cancer
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted fluorescence microscope under 100 X oil immersion. .. SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing five copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository [ ] and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 h, followed by 30 μM cisplatin for 24 h, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: A tight balance of Karyopherin β1 expression is required in cervical cancer cells
    Article Snippet: To assay for NFAT luciferase activity, cells were transfected with 50 ng GFP-NFAT (Addgene plasmid #24219; gift of Jerry Crabtree [ ]), 50 ng NFAT-luciferase (Addgene plasmid #10959; gift of Toren Finkel [ ]), and 5 ng pRL-TK (encoding Renilla luciferase; Promega, USA). .. To assay for AP-1, NFκB p65 and p53 luciferase activity, cells were transfected with either 100 ng AP1-luciferase reporter construct (containing four copies of the AP-1 binding site; gift of Michael Birrer [ ]), 100 ng NFκB p65 luciferase reporter construct (containing 5 copies of the p65 binding site; Promega, USA) or 100 ng p53-luciferase reporter construct (Addgene plasmid #16442; gift of Bert Vogelstein [ ]), as well as 5 ng pRL-TK. .. For NFκB p65 luciferase assays, cells were stimulated with 0.5 μM PMA (Sigma-Aldrich, USA) for 3 h, at 24 h post transfection.

    Expressing:

    Article Title: HIPK2 C-terminal domain inhibits NF-κB signaling and renal inflammation in kidney injury.
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, #SL100688). .. Luciferase reporter gene assays HEK293T cells were co-transfected with pcDNA4B or HIPK2CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Dr. Huabao Xiong (28), and SBE-luc and p53-luc were obtained from Addgene). ..

    Article Title: HIPK2 C-terminal domain inhibits NF- κ B signaling and renal inflammation in kidney injury
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, SL100688). .. HEK293T cells were cotransfected with pcDNA4B or HIPK2-CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Huabao Xiong [Department of Medicine, Precision Immunology Institute, Icahn School of Medicine at Mount Sinai], ref. , and SBE-luc and p53-luc were obtained from Addgene). ..

    Plasmid Preparation:

    Article Title: HIPK2 C-terminal domain inhibits NF-κB signaling and renal inflammation in kidney injury.
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, #SL100688). .. Luciferase reporter gene assays HEK293T cells were co-transfected with pcDNA4B or HIPK2CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Dr. Huabao Xiong (28), and SBE-luc and p53-luc were obtained from Addgene). ..

    Article Title: HIPK2 C-terminal domain inhibits NF- κ B signaling and renal inflammation in kidney injury
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, SL100688). .. HEK293T cells were cotransfected with pcDNA4B or HIPK2-CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Huabao Xiong [Department of Medicine, Precision Immunology Institute, Icahn School of Medicine at Mount Sinai], ref. , and SBE-luc and p53-luc were obtained from Addgene). ..

    Article Title: Inhibition of Kpn β 1 Mediated Nuclear Import Enhances Cisplatin Chemosensitivity in Cervical Cancer.
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted uorescence microscope under 100 X oil immersion. .. Page 6/23 Luciferase reporter assay SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing ve copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository(26) and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 hours, followed by 30 μM cisplatin for 24 hours, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: Inhibition of Kpnβ1 mediated nuclear import enhances cisplatin chemosensitivity in cervical cancer
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted fluorescence microscope under 100 X oil immersion. .. SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing five copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository [ ] and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 h, followed by 30 μM cisplatin for 24 h, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: A tight balance of Karyopherin β1 expression is required in cervical cancer cells
    Article Snippet: To assay for NFAT luciferase activity, cells were transfected with 50 ng GFP-NFAT (Addgene plasmid #24219; gift of Jerry Crabtree [ ]), 50 ng NFAT-luciferase (Addgene plasmid #10959; gift of Toren Finkel [ ]), and 5 ng pRL-TK (encoding Renilla luciferase; Promega, USA). .. To assay for AP-1, NFκB p65 and p53 luciferase activity, cells were transfected with either 100 ng AP1-luciferase reporter construct (containing four copies of the AP-1 binding site; gift of Michael Birrer [ ]), 100 ng NFκB p65 luciferase reporter construct (containing 5 copies of the p65 binding site; Promega, USA) or 100 ng p53-luciferase reporter construct (Addgene plasmid #16442; gift of Bert Vogelstein [ ]), as well as 5 ng pRL-TK. .. For NFκB p65 luciferase assays, cells were stimulated with 0.5 μM PMA (Sigma-Aldrich, USA) for 3 h, at 24 h post transfection.

    Transfection:

    Article Title: HIPK2 C-terminal domain inhibits NF-κB signaling and renal inflammation in kidney injury.
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, #SL100688). .. Luciferase reporter gene assays HEK293T cells were co-transfected with pcDNA4B or HIPK2CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Dr. Huabao Xiong (28), and SBE-luc and p53-luc were obtained from Addgene). ..

    Article Title: HIPK2 C-terminal domain inhibits NF- κ B signaling and renal inflammation in kidney injury
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, SL100688). .. HEK293T cells were cotransfected with pcDNA4B or HIPK2-CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Huabao Xiong [Department of Medicine, Precision Immunology Institute, Icahn School of Medicine at Mount Sinai], ref. , and SBE-luc and p53-luc were obtained from Addgene). ..

    Article Title: Inhibition of Kpn β 1 Mediated Nuclear Import Enhances Cisplatin Chemosensitivity in Cervical Cancer.
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted uorescence microscope under 100 X oil immersion. .. Page 6/23 Luciferase reporter assay SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing ve copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository(26) and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 hours, followed by 30 μM cisplatin for 24 hours, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: Inhibition of Kpnβ1 mediated nuclear import enhances cisplatin chemosensitivity in cervical cancer
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted fluorescence microscope under 100 X oil immersion. .. SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing five copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository [ ] and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 h, followed by 30 μM cisplatin for 24 h, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: A tight balance of Karyopherin β1 expression is required in cervical cancer cells
    Article Snippet: To assay for NFAT luciferase activity, cells were transfected with 50 ng GFP-NFAT (Addgene plasmid #24219; gift of Jerry Crabtree [ ]), 50 ng NFAT-luciferase (Addgene plasmid #10959; gift of Toren Finkel [ ]), and 5 ng pRL-TK (encoding Renilla luciferase; Promega, USA). .. To assay for AP-1, NFκB p65 and p53 luciferase activity, cells were transfected with either 100 ng AP1-luciferase reporter construct (containing four copies of the AP-1 binding site; gift of Michael Birrer [ ]), 100 ng NFκB p65 luciferase reporter construct (containing 5 copies of the p65 binding site; Promega, USA) or 100 ng p53-luciferase reporter construct (Addgene plasmid #16442; gift of Bert Vogelstein [ ]), as well as 5 ng pRL-TK. .. For NFκB p65 luciferase assays, cells were stimulated with 0.5 μM PMA (Sigma-Aldrich, USA) for 3 h, at 24 h post transfection.

    Construct:

    Article Title: HIPK2 C-terminal domain inhibits NF-κB signaling and renal inflammation in kidney injury.
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, #SL100688). .. Luciferase reporter gene assays HEK293T cells were co-transfected with pcDNA4B or HIPK2CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Dr. Huabao Xiong (28), and SBE-luc and p53-luc were obtained from Addgene). ..

    Article Title: HIPK2 C-terminal domain inhibits NF- κ B signaling and renal inflammation in kidney injury
    Article Snippet: For transient transfection, HEK293T cells were transfected using PolyJet reagent (SignaGen, SL100688). .. HEK293T cells were cotransfected with pcDNA4B or HIPK2-CT, as well as Renilla expression vector to normalize transfection efficiency, and p65 luciferase reporter construct or Smad3-binding element luciferase reporter construct or p53 luciferase reporter construct (p65-luc construct was a gift from the laboratory of Huabao Xiong [Department of Medicine, Precision Immunology Institute, Icahn School of Medicine at Mount Sinai], ref. , and SBE-luc and p53-luc were obtained from Addgene). ..

    Article Title: Inhibition of Kpn β 1 Mediated Nuclear Import Enhances Cisplatin Chemosensitivity in Cervical Cancer.
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted uorescence microscope under 100 X oil immersion. .. Page 6/23 Luciferase reporter assay SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing ve copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository(26) and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 hours, followed by 30 μM cisplatin for 24 hours, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: Inhibition of Kpnβ1 mediated nuclear import enhances cisplatin chemosensitivity in cervical cancer
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted fluorescence microscope under 100 X oil immersion. .. SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing five copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository [ ] and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 h, followed by 30 μM cisplatin for 24 h, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: A tight balance of Karyopherin β1 expression is required in cervical cancer cells
    Article Snippet: To assay for NFAT luciferase activity, cells were transfected with 50 ng GFP-NFAT (Addgene plasmid #24219; gift of Jerry Crabtree [ ]), 50 ng NFAT-luciferase (Addgene plasmid #10959; gift of Toren Finkel [ ]), and 5 ng pRL-TK (encoding Renilla luciferase; Promega, USA). .. To assay for AP-1, NFκB p65 and p53 luciferase activity, cells were transfected with either 100 ng AP1-luciferase reporter construct (containing four copies of the AP-1 binding site; gift of Michael Birrer [ ]), 100 ng NFκB p65 luciferase reporter construct (containing 5 copies of the p65 binding site; Promega, USA) or 100 ng p53-luciferase reporter construct (Addgene plasmid #16442; gift of Bert Vogelstein [ ]), as well as 5 ng pRL-TK. .. For NFκB p65 luciferase assays, cells were stimulated with 0.5 μM PMA (Sigma-Aldrich, USA) for 3 h, at 24 h post transfection.

    Reporter Assay:

    Article Title: Inhibition of Kpn β 1 Mediated Nuclear Import Enhances Cisplatin Chemosensitivity in Cervical Cancer.
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted uorescence microscope under 100 X oil immersion. .. Page 6/23 Luciferase reporter assay SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing ve copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository(26) and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 hours, followed by 30 μM cisplatin for 24 hours, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Binding Assay:

    Article Title: Inhibition of Kpn β 1 Mediated Nuclear Import Enhances Cisplatin Chemosensitivity in Cervical Cancer.
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted uorescence microscope under 100 X oil immersion. .. Page 6/23 Luciferase reporter assay SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing ve copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository(26) and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 hours, followed by 30 μM cisplatin for 24 hours, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: Inhibition of Kpnβ1 mediated nuclear import enhances cisplatin chemosensitivity in cervical cancer
    Article Snippet: Cell nuclei were counterstained with 0.5 μg/mL DAPI, and images captured using the Zeiss inverted fluorescence microscope under 100 X oil immersion. .. SiHa cells were transfected with 100 ng p65-luciferase reporter construct (containing five copies of the p65-binding site, Promega) or 200 ng p53-luciferase reporter construct (containing thirteen wildtype p53 binding sites, Addgene plasmid #16442, Addgene Plasmid Repository [ ] and 10 ng pRL-TK (encoding Renilla luciferase, Promega), using Genecellin transfection reagent (Celtic Molecular Diagnostics). .. The following day cells were treated with 5 μM INI-43 for 2 h, followed by 30 μM cisplatin for 24 h, and luciferase activity assayed using the Dual-Luciferase Report assay system (Promega), according to the manufacturer’s instructions.

    Article Title: A tight balance of Karyopherin β1 expression is required in cervical cancer cells
    Article Snippet: To assay for NFAT luciferase activity, cells were transfected with 50 ng GFP-NFAT (Addgene plasmid #24219; gift of Jerry Crabtree [ ]), 50 ng NFAT-luciferase (Addgene plasmid #10959; gift of Toren Finkel [ ]), and 5 ng pRL-TK (encoding Renilla luciferase; Promega, USA). .. To assay for AP-1, NFκB p65 and p53 luciferase activity, cells were transfected with either 100 ng AP1-luciferase reporter construct (containing four copies of the AP-1 binding site; gift of Michael Birrer [ ]), 100 ng NFκB p65 luciferase reporter construct (containing 5 copies of the p65 binding site; Promega, USA) or 100 ng p53-luciferase reporter construct (Addgene plasmid #16442; gift of Bert Vogelstein [ ]), as well as 5 ng pRL-TK. .. For NFκB p65 luciferase assays, cells were stimulated with 0.5 μM PMA (Sigma-Aldrich, USA) for 3 h, at 24 h post transfection.

    Activity Assay:

    Article Title: A tight balance of Karyopherin β1 expression is required in cervical cancer cells
    Article Snippet: To assay for NFAT luciferase activity, cells were transfected with 50 ng GFP-NFAT (Addgene plasmid #24219; gift of Jerry Crabtree [ ]), 50 ng NFAT-luciferase (Addgene plasmid #10959; gift of Toren Finkel [ ]), and 5 ng pRL-TK (encoding Renilla luciferase; Promega, USA). .. To assay for AP-1, NFκB p65 and p53 luciferase activity, cells were transfected with either 100 ng AP1-luciferase reporter construct (containing four copies of the AP-1 binding site; gift of Michael Birrer [ ]), 100 ng NFκB p65 luciferase reporter construct (containing 5 copies of the p65 binding site; Promega, USA) or 100 ng p53-luciferase reporter construct (Addgene plasmid #16442; gift of Bert Vogelstein [ ]), as well as 5 ng pRL-TK. .. For NFκB p65 luciferase assays, cells were stimulated with 0.5 μM PMA (Sigma-Aldrich, USA) for 3 h, at 24 h post transfection.



    Similar Products

    p53 luciferase reporter construct  (Addgene inc)
    92
    Addgene inc p53 luciferase reporter construct
    P53 Luciferase Reporter Construct, supplied by Addgene inc, 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/product/p53+luciferase+reporter+construct/GFP-p53+(NES-)+(Plasmid+%2312092)/pm38512421-113-34-58
    Average 92 stars, based on 1 article reviews
    p53 luciferase reporter construct - by Bioz Stars, 2026-10
    92/100 stars
    		  Buy from Supplier
    luciferase reporter construct with p53-responsible element  (Promega)
    90
    Promega luciferase reporter construct with p53-responsible element
    Luciferase Reporter Construct With P53 Responsible Element, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p53+luciferase+reporter+construct/luciferase+reporter+construct+with+p53+responsible+element/pm35472819-105-5-10
    Average 90 stars, based on 1 article reviews
    luciferase reporter construct with p53-responsible element - by Bioz Stars, 2026-10
    90/100 stars
    		  Buy from Supplier
    luciferase reporter construct pgl4.38 [luc2p/p53 re/hygro]  (Promega)
    90
    Promega luciferase reporter construct pgl4.38 [luc2p/p53 re/hygro]
    a Colony formation assay: A375 and MEL-JUSO cells were treated with 20 µM roscovitine or 500 nM flavopiridol for 24 h, washed, and further cultivated in a drug-free growth medium. Cells were fixed and stained with 6.0 % glutaraldehyde + 0.5 % crystal violet solution. b Activation of <t>p53</t> by roscovitine and flavopiridol: A375 and MEL-JUSO cells stably transfected with a p53 transcriptional activity luciferase reporter construct were treated with the indicated concentrations of roscovitine and flavopiridol for 24 h, lysed, and luciferase activity in lysates was determined. Relative p53 transcriptional activity was calculated. The values represent the mean ± SD; N = 3; * P < 0.05. c Roscovitine and flavopiridol downregulate MDM4 at lower concentrations than MDM2, concurrent MDM4 and MDM2 downregulation leads to stabilization of p53. Western blot analysis of protein lysates of A375 and MEL-JUSO cells after 24-h treatment with CDK inhibitors. The levels of PCNA protein served as a loading control.
    Luciferase Reporter Construct Pgl4.38 [Luc2p/P53 Re/Hygro], supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p53+luciferase+reporter+construct/pgl3+basic/pmc07494941-228-19-26
    Average 90 stars, based on 1 article reviews
    luciferase reporter construct pgl4.38 [luc2p/p53 re/hygro] - by Bioz Stars, 2026-10
    90/100 stars
    		  Buy from Supplier
    p53-responsive luciferase reporter construct pgl4.38[luc2p/p53]  (Tivity Health Inc)
    90
    Tivity Health Inc p53-responsive luciferase reporter construct pgl4.38[luc2p/p53]
    a Colony formation assay: A375 and MEL-JUSO cells were treated with 20 µM roscovitine or 500 nM flavopiridol for 24 h, washed, and further cultivated in a drug-free growth medium. Cells were fixed and stained with 6.0 % glutaraldehyde + 0.5 % crystal violet solution. b Activation of <t>p53</t> by roscovitine and flavopiridol: A375 and MEL-JUSO cells stably transfected with a p53 transcriptional activity luciferase reporter construct were treated with the indicated concentrations of roscovitine and flavopiridol for 24 h, lysed, and luciferase activity in lysates was determined. Relative p53 transcriptional activity was calculated. The values represent the mean ± SD; N = 3; * P < 0.05. c Roscovitine and flavopiridol downregulate MDM4 at lower concentrations than MDM2, concurrent MDM4 and MDM2 downregulation leads to stabilization of p53. Western blot analysis of protein lysates of A375 and MEL-JUSO cells after 24-h treatment with CDK inhibitors. The levels of PCNA protein served as a loading control.
    P53 Responsive Luciferase Reporter Construct Pgl4.38[Luc2p/P53], supplied by Tivity Health Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p53+luciferase+reporter+construct/p53+responsive+luciferase+reporter+construct+pgl4+38+luc2p+p53+/pm31227395-128-34-39
    Average 90 stars, based on 1 article reviews
    p53-responsive luciferase reporter construct pgl4.38[luc2p/p53] - by Bioz Stars, 2026-10
    90/100 stars
    		  Buy from Supplier
    p53-luciferase reporter construct  (Qiagen)
    90
    Qiagen p53-luciferase reporter construct
    MDM2 inhibitors induce a senescence-like state. ( A , B ) HCA2 fibroblasts were treated using the indicated concentrations of MI-63 ( A ) or nutlin-3a ( B ). <t>p53</t> and p21 levels were analyzed by western blotting. Actin levels served as a loading control. ( C ) IMR-90 fibroblasts were transduced with a <t>p53</t> <t>luciferase</t> reporter and treated with MI-63 or nutlin-3a. Extracts were prepared and analyzed by luminometry. ( D , E ) HCA2 cells were treated with MI-63 or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in ( D ) and percentage of positive cells is shown in ( E ). ( F , G ) HCA2 cells were treated with nutlin-3a or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in ( F ) and percentage of positive cells is shown in ( G ). ( H-I ) HCA2 cells were irradiated (IR) or treated with MI-63 or nutlin-3a, and immunostained for γH2AX. Representative images are shown in ( H ) and percentage of cells with >3 γH2AX nuclear foci is shown in ( I ). Data are representative of two independent experiments. ***p < 0.001 by 1-way ANOVA.
    P53 Luciferase Reporter Construct, supplied by Qiagen, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p53+luciferase+reporter+construct/p53+response+luciferase+construct/pmc05799282-173-11-14
    Average 90 stars, based on 1 article reviews
    p53-luciferase reporter construct - by Bioz Stars, 2026-10
    90/100 stars
    		  Buy from Supplier
    p53 reporter construct coding for firefly luciferase  (Addgene inc)
    90
    Addgene inc p53 reporter construct coding for firefly luciferase
    MDM2 inhibitors induce a senescence-like state. ( A , B ) HCA2 fibroblasts were treated using the indicated concentrations of MI-63 ( A ) or nutlin-3a ( B ). <t>p53</t> and p21 levels were analyzed by western blotting. Actin levels served as a loading control. ( C ) IMR-90 fibroblasts were transduced with a <t>p53</t> <t>luciferase</t> reporter and treated with MI-63 or nutlin-3a. Extracts were prepared and analyzed by luminometry. ( D , E ) HCA2 cells were treated with MI-63 or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in ( D ) and percentage of positive cells is shown in ( E ). ( F , G ) HCA2 cells were treated with nutlin-3a or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in ( F ) and percentage of positive cells is shown in ( G ). ( H-I ) HCA2 cells were irradiated (IR) or treated with MI-63 or nutlin-3a, and immunostained for γH2AX. Representative images are shown in ( H ) and percentage of cells with >3 γH2AX nuclear foci is shown in ( I ). Data are representative of two independent experiments. ***p < 0.001 by 1-way ANOVA.
    P53 Reporter Construct Coding For Firefly Luciferase, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p53+luciferase+reporter+construct/p53+reporter+plasmid/pm26972256-42-7-11
    Average 90 stars, based on 1 article reviews
    p53 reporter construct coding for firefly luciferase - by Bioz Stars, 2026-10
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    a Colony formation assay: A375 and MEL-JUSO cells were treated with 20 µM roscovitine or 500 nM flavopiridol for 24 h, washed, and further cultivated in a drug-free growth medium. Cells were fixed and stained with 6.0 % glutaraldehyde + 0.5 % crystal violet solution. b Activation of p53 by roscovitine and flavopiridol: A375 and MEL-JUSO cells stably transfected with a p53 transcriptional activity luciferase reporter construct were treated with the indicated concentrations of roscovitine and flavopiridol for 24 h, lysed, and luciferase activity in lysates was determined. Relative p53 transcriptional activity was calculated. The values represent the mean ± SD; N = 3; * P < 0.05. c Roscovitine and flavopiridol downregulate MDM4 at lower concentrations than MDM2, concurrent MDM4 and MDM2 downregulation leads to stabilization of p53. Western blot analysis of protein lysates of A375 and MEL-JUSO cells after 24-h treatment with CDK inhibitors. The levels of PCNA protein served as a loading control.

    Journal: Cell Death & Disease

    Article Title: CDK9 activity is critical for maintaining MDM4 overexpression in tumor cells

    doi: 10.1038/s41419-020-02971-3

    Figure Lengend Snippet: a Colony formation assay: A375 and MEL-JUSO cells were treated with 20 µM roscovitine or 500 nM flavopiridol for 24 h, washed, and further cultivated in a drug-free growth medium. Cells were fixed and stained with 6.0 % glutaraldehyde + 0.5 % crystal violet solution. b Activation of p53 by roscovitine and flavopiridol: A375 and MEL-JUSO cells stably transfected with a p53 transcriptional activity luciferase reporter construct were treated with the indicated concentrations of roscovitine and flavopiridol for 24 h, lysed, and luciferase activity in lysates was determined. Relative p53 transcriptional activity was calculated. The values represent the mean ± SD; N = 3; * P < 0.05. c Roscovitine and flavopiridol downregulate MDM4 at lower concentrations than MDM2, concurrent MDM4 and MDM2 downregulation leads to stabilization of p53. Western blot analysis of protein lysates of A375 and MEL-JUSO cells after 24-h treatment with CDK inhibitors. The levels of PCNA protein served as a loading control.

    Article Snippet: Wild-type p53 melanoma cell lines A375 and MEL-JUSO and the hESC cell line CCTL14 were stably transfected with the p53-responsive luciferase reporter construct pGL4.38 [luc2P/p53 RE/Hygro] (Promega).

    Techniques: Colony Assay, Staining, Activation Assay, Stable Transfection, Transfection, Activity Assay, Luciferase, Construct, Western Blot, Control

    a Activation of p53-dependent transcription by CDK9 inhibitor atuveciclib and CDK1 inhibitor RO3306. A375 cells expressing luciferase under the control of a p53-responsive promoter were treated with specific CDK inhibitors for 24 h. Relative p53 activity in cell lysates was determined. The values represent the mean ± SD; N = 3; * P < 0.05; ** P < 0.01. b Inhibitors of transcriptional CDKs THZ1, THZ531, and atuveciclib inhibit MDM4 expression and stabilize p53, with minimal impact on MDM2 levels. Western blot analysis of A375 cell lysates after 24-h treatment with the indicated concentrations of specific CDK inhibitors. c Time-course experiment with CDK9 inhibitor atuveciclib. A375 cells were treated with atuveciclib for 1, 3, 6, and 16 h. Proteins of interest were visualized by western blotting. d CDK9 knockdown downregulates MDM4 levels. Western blot analysis of A375 response to siRNA-mediated CDK9 knockdown. Commercially available non-targeting siRNAs (control siRNA 1) and GFP-targeting siRNAs (control siRNA 2 and 3) were used as controls. e CDK9 PROTAC THAL-SNS-032. The dual affinity molecule binds E3 ubiquitin ligase CRBN and induces selective degradation of the CDK9 protein. f Selective knockdown of CDK9 by THAL-SNS-032 induces a decrease in MDM4 levels. Western blot analysis of A375 cell lysates after 1, 6, and 24-h treatment with the indicated concentrations of THAL-SNS-032.

    Journal: Cell Death & Disease

    Article Title: CDK9 activity is critical for maintaining MDM4 overexpression in tumor cells

    doi: 10.1038/s41419-020-02971-3

    Figure Lengend Snippet: a Activation of p53-dependent transcription by CDK9 inhibitor atuveciclib and CDK1 inhibitor RO3306. A375 cells expressing luciferase under the control of a p53-responsive promoter were treated with specific CDK inhibitors for 24 h. Relative p53 activity in cell lysates was determined. The values represent the mean ± SD; N = 3; * P < 0.05; ** P < 0.01. b Inhibitors of transcriptional CDKs THZ1, THZ531, and atuveciclib inhibit MDM4 expression and stabilize p53, with minimal impact on MDM2 levels. Western blot analysis of A375 cell lysates after 24-h treatment with the indicated concentrations of specific CDK inhibitors. c Time-course experiment with CDK9 inhibitor atuveciclib. A375 cells were treated with atuveciclib for 1, 3, 6, and 16 h. Proteins of interest were visualized by western blotting. d CDK9 knockdown downregulates MDM4 levels. Western blot analysis of A375 response to siRNA-mediated CDK9 knockdown. Commercially available non-targeting siRNAs (control siRNA 1) and GFP-targeting siRNAs (control siRNA 2 and 3) were used as controls. e CDK9 PROTAC THAL-SNS-032. The dual affinity molecule binds E3 ubiquitin ligase CRBN and induces selective degradation of the CDK9 protein. f Selective knockdown of CDK9 by THAL-SNS-032 induces a decrease in MDM4 levels. Western blot analysis of A375 cell lysates after 1, 6, and 24-h treatment with the indicated concentrations of THAL-SNS-032.

    Article Snippet: Wild-type p53 melanoma cell lines A375 and MEL-JUSO and the hESC cell line CCTL14 were stably transfected with the p53-responsive luciferase reporter construct pGL4.38 [luc2P/p53 RE/Hygro] (Promega).

    Techniques: Activation Assay, Expressing, Luciferase, Control, Activity Assay, Western Blot, Knockdown, Ubiquitin Proteomics

    a MDM4 gene expression analysis by qRT-PCR. Six-hour treatment with dinaciclib and atuveciclib was used to inhibit CDK9-dependent transcription in A375 cells. Total RNA was isolated using RNA Blue reagent, reverse transcribed, and real-time quantitative PCR was performed in triplicates. The values represent the mean ± SD; N = 4; * P < 0.05. b mNET-Seq analysis of RNA polymerase II position along the MDM4 gene. Raji B cells harboring an analog sensitive CDK9 mutation were treated with adenine analog 1-NA-PP1 (CDK9as inhibited) or DMSO (control), and RNA polymerase II occupancy within the MDM4 gene region between control and CDK9-inhibited cells was compared. c Western blot analysis of MDM4, MDM2, p53, and p21 protein levels in A375 cell lysates after 24-hour treatment with the indicated concentrations of CDK9 inhibitor atuveciclib. PCNA served as a loading control. d Expression analysis of p53 target genes p21 and PUMA by qRT-PCR. A375 cells were treated with CDK9 inhibitor atuveciclib for 14 h before harvesting. Total RNA was isolated using RNA Blue reagent, reverse transcribed, and real-time quantitative PCR was performed in triplicates. The values represent the mean ± SD; N = 3; * P < 0.05; ** P < 0.01.

    Journal: Cell Death & Disease

    Article Title: CDK9 activity is critical for maintaining MDM4 overexpression in tumor cells

    doi: 10.1038/s41419-020-02971-3

    Figure Lengend Snippet: a MDM4 gene expression analysis by qRT-PCR. Six-hour treatment with dinaciclib and atuveciclib was used to inhibit CDK9-dependent transcription in A375 cells. Total RNA was isolated using RNA Blue reagent, reverse transcribed, and real-time quantitative PCR was performed in triplicates. The values represent the mean ± SD; N = 4; * P < 0.05. b mNET-Seq analysis of RNA polymerase II position along the MDM4 gene. Raji B cells harboring an analog sensitive CDK9 mutation were treated with adenine analog 1-NA-PP1 (CDK9as inhibited) or DMSO (control), and RNA polymerase II occupancy within the MDM4 gene region between control and CDK9-inhibited cells was compared. c Western blot analysis of MDM4, MDM2, p53, and p21 protein levels in A375 cell lysates after 24-hour treatment with the indicated concentrations of CDK9 inhibitor atuveciclib. PCNA served as a loading control. d Expression analysis of p53 target genes p21 and PUMA by qRT-PCR. A375 cells were treated with CDK9 inhibitor atuveciclib for 14 h before harvesting. Total RNA was isolated using RNA Blue reagent, reverse transcribed, and real-time quantitative PCR was performed in triplicates. The values represent the mean ± SD; N = 3; * P < 0.05; ** P < 0.01.

    Article Snippet: Wild-type p53 melanoma cell lines A375 and MEL-JUSO and the hESC cell line CCTL14 were stably transfected with the p53-responsive luciferase reporter construct pGL4.38 [luc2P/p53 RE/Hygro] (Promega).

    Techniques: Gene Expression, Quantitative RT-PCR, Isolation, Reverse Transcription, Real-time Polymerase Chain Reaction, Mutagenesis, Control, Western Blot, Expressing

    a Activation of p53 by atuveciclib combined with nutlin-3a treatment. To avoid a complete transcriptional block and thus interference with the luciferase reporter system, A375 cells were pretreated with atuveciclib for 16 h, washed, and treated with nutlin-3a for 10 h. DMSO pretreatment and treatment was used for control = DMSO + DMSO; ctrl = untreated control. The values are relative to the DMSO + nutlin-3a sample and represent the mean ± SD; N = 3; * P < 0.05; *** P < 0.001. b Western blot analysis of cell lysates after 24-h co-treatment with atuveciclib and nutlin-3a. c Effect of the drug combination on melanoma cell viability. Propidium iodide viability assay was performed after a 48-h co-treatment with atuveciclib and nutlin-3a in A375 and MEL-JUSO cells. The values represent the mean ± SD; N = 3; ** P < 0.01; *** P < 0.001.

    Journal: Cell Death & Disease

    Article Title: CDK9 activity is critical for maintaining MDM4 overexpression in tumor cells

    doi: 10.1038/s41419-020-02971-3

    Figure Lengend Snippet: a Activation of p53 by atuveciclib combined with nutlin-3a treatment. To avoid a complete transcriptional block and thus interference with the luciferase reporter system, A375 cells were pretreated with atuveciclib for 16 h, washed, and treated with nutlin-3a for 10 h. DMSO pretreatment and treatment was used for control = DMSO + DMSO; ctrl = untreated control. The values are relative to the DMSO + nutlin-3a sample and represent the mean ± SD; N = 3; * P < 0.05; *** P < 0.001. b Western blot analysis of cell lysates after 24-h co-treatment with atuveciclib and nutlin-3a. c Effect of the drug combination on melanoma cell viability. Propidium iodide viability assay was performed after a 48-h co-treatment with atuveciclib and nutlin-3a in A375 and MEL-JUSO cells. The values represent the mean ± SD; N = 3; ** P < 0.01; *** P < 0.001.

    Article Snippet: Wild-type p53 melanoma cell lines A375 and MEL-JUSO and the hESC cell line CCTL14 were stably transfected with the p53-responsive luciferase reporter construct pGL4.38 [luc2P/p53 RE/Hygro] (Promega).

    Techniques: Activation Assay, Blocking Assay, Luciferase, Control, Western Blot, Viability Assay

    a CDK9 inhibition leads to MDM4 downregulation in hESCs. Western blot analysis of CCTL14 and CCTL12 cell lysates after 24-h treatment with selected CDK inhibitors. b Selective knockdown of CDK9 by THAL-SNS-032 PROTAC promotes MDM4 depletion in hESCs. Western blot analysis of CCTL14 lysates after 16-h THAL-SNS-032 treatment. c MDM4/MDM2 knockdown does not affect p53 levels in hESCs. Western blot and p53 transcriptional activity analysis in hESCs upon siRNA-mediated knockdown of MDM2 and MDM4. DNA-damaging agent doxorubicin served as a positive control. The p53 activity values are relative to control and error bars represent the SD; N = 3; double asterisk denotes P < 0.01.

    Journal: Cell Death & Disease

    Article Title: CDK9 activity is critical for maintaining MDM4 overexpression in tumor cells

    doi: 10.1038/s41419-020-02971-3

    Figure Lengend Snippet: a CDK9 inhibition leads to MDM4 downregulation in hESCs. Western blot analysis of CCTL14 and CCTL12 cell lysates after 24-h treatment with selected CDK inhibitors. b Selective knockdown of CDK9 by THAL-SNS-032 PROTAC promotes MDM4 depletion in hESCs. Western blot analysis of CCTL14 lysates after 16-h THAL-SNS-032 treatment. c MDM4/MDM2 knockdown does not affect p53 levels in hESCs. Western blot and p53 transcriptional activity analysis in hESCs upon siRNA-mediated knockdown of MDM2 and MDM4. DNA-damaging agent doxorubicin served as a positive control. The p53 activity values are relative to control and error bars represent the SD; N = 3; double asterisk denotes P < 0.01.

    Article Snippet: Wild-type p53 melanoma cell lines A375 and MEL-JUSO and the hESC cell line CCTL14 were stably transfected with the p53-responsive luciferase reporter construct pGL4.38 [luc2P/p53 RE/Hygro] (Promega).

    Techniques: Inhibition, Western Blot, Knockdown, Activity Assay, Positive Control, Control

    MDM2 inhibitors induce a senescence-like state. ( A , B ) HCA2 fibroblasts were treated using the indicated concentrations of MI-63 ( A ) or nutlin-3a ( B ). p53 and p21 levels were analyzed by western blotting. Actin levels served as a loading control. ( C ) IMR-90 fibroblasts were transduced with a p53 luciferase reporter and treated with MI-63 or nutlin-3a. Extracts were prepared and analyzed by luminometry. ( D , E ) HCA2 cells were treated with MI-63 or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in ( D ) and percentage of positive cells is shown in ( E ). ( F , G ) HCA2 cells were treated with nutlin-3a or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in ( F ) and percentage of positive cells is shown in ( G ). ( H-I ) HCA2 cells were irradiated (IR) or treated with MI-63 or nutlin-3a, and immunostained for γH2AX. Representative images are shown in ( H ) and percentage of cells with >3 γH2AX nuclear foci is shown in ( I ). Data are representative of two independent experiments. ***p < 0.001 by 1-way ANOVA.

    Journal: Scientific Reports

    Article Title: Small-molecule MDM2 antagonists attenuate the senescence-associated secretory phenotype

    doi: 10.1038/s41598-018-20000-4

    Figure Lengend Snippet: MDM2 inhibitors induce a senescence-like state. ( A , B ) HCA2 fibroblasts were treated using the indicated concentrations of MI-63 ( A ) or nutlin-3a ( B ). p53 and p21 levels were analyzed by western blotting. Actin levels served as a loading control. ( C ) IMR-90 fibroblasts were transduced with a p53 luciferase reporter and treated with MI-63 or nutlin-3a. Extracts were prepared and analyzed by luminometry. ( D , E ) HCA2 cells were treated with MI-63 or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in ( D ) and percentage of positive cells is shown in ( E ). ( F , G ) HCA2 cells were treated with nutlin-3a or induced to senesce by 10 Gy IR. SA-β-gal staining is shown in ( F ) and percentage of positive cells is shown in ( G ). ( H-I ) HCA2 cells were irradiated (IR) or treated with MI-63 or nutlin-3a, and immunostained for γH2AX. Representative images are shown in ( H ) and percentage of cells with >3 γH2AX nuclear foci is shown in ( I ). Data are representative of two independent experiments. ***p < 0.001 by 1-way ANOVA.

    Article Snippet: Cells were co-transduced with a lentiviral NF-κB luciferase reporter construct or p53-luciferase reporter construct (SA Biosciences), and a constitutively expressed renilla luciferase construct (SA Biosciences).

    Techniques: Western Blot, Transduction, Luciferase, Staining, Irradiation

    MDM2 antagonist attenuates the SASP. ( A ) IL-6 ELISA using conditioned media (CM) from HCA2 and IMR-90 fibroblasts that were induced to senesce by IR and treated with DMSO or nutlin-3a at two concentrations. ( B ) IL-6 in CM from IMR-90 cells (mock or IR) after treatment with DMSO, rapamycin (RAPA) or MI-63. ( C , D ) IMR-90 cells were treated for 24 h with the chemotherapeutic agents doxorubicin (DOXO) ( C ), taxol or cisplatin ( D ), and IL-6 secretion measured by ELISA in cells cultured with MI-63 ( C ) or nutlin-3a ( D ) for 24 h following removal of the drugs. ( E ) Heat map indicating gene expression of several SASP factors in IMR-90 fibroblasts (mock or IR) treated DMSO or nutlin-3a. ( F , G ) Relative expression of ( F ) IL-1α and ( G ) IL-1β in control or irradiated (IR) fibroblasts cultured with nutlin-3a or DMSO. ( H ) HCA2 cells were transduced with a lentiviral NF-κB luciferase reporter construct, and luciferase activity in DMSO- or nutlin-3a-treated (mock or IR) cells was measured. ( I ) HCA2 fibroblasts were transduced with lentiviruses expressing either control (shGFP) or p53-targeted shRNAs (shp53) or a p53 genetic suppressor element (GSE22) and treated with DMSO or nutlin-3a for 24 h following mock or X-irradiation (IR). IL-6 secretion was measured by ELISA. Data are representative of three independent experiments. *p < 0.05, ***p < 0.001 by 2-way ANOVA.

    Journal: Scientific Reports

    Article Title: Small-molecule MDM2 antagonists attenuate the senescence-associated secretory phenotype

    doi: 10.1038/s41598-018-20000-4

    Figure Lengend Snippet: MDM2 antagonist attenuates the SASP. ( A ) IL-6 ELISA using conditioned media (CM) from HCA2 and IMR-90 fibroblasts that were induced to senesce by IR and treated with DMSO or nutlin-3a at two concentrations. ( B ) IL-6 in CM from IMR-90 cells (mock or IR) after treatment with DMSO, rapamycin (RAPA) or MI-63. ( C , D ) IMR-90 cells were treated for 24 h with the chemotherapeutic agents doxorubicin (DOXO) ( C ), taxol or cisplatin ( D ), and IL-6 secretion measured by ELISA in cells cultured with MI-63 ( C ) or nutlin-3a ( D ) for 24 h following removal of the drugs. ( E ) Heat map indicating gene expression of several SASP factors in IMR-90 fibroblasts (mock or IR) treated DMSO or nutlin-3a. ( F , G ) Relative expression of ( F ) IL-1α and ( G ) IL-1β in control or irradiated (IR) fibroblasts cultured with nutlin-3a or DMSO. ( H ) HCA2 cells were transduced with a lentiviral NF-κB luciferase reporter construct, and luciferase activity in DMSO- or nutlin-3a-treated (mock or IR) cells was measured. ( I ) HCA2 fibroblasts were transduced with lentiviruses expressing either control (shGFP) or p53-targeted shRNAs (shp53) or a p53 genetic suppressor element (GSE22) and treated with DMSO or nutlin-3a for 24 h following mock or X-irradiation (IR). IL-6 secretion was measured by ELISA. Data are representative of three independent experiments. *p < 0.05, ***p < 0.001 by 2-way ANOVA.

    Article Snippet: Cells were co-transduced with a lentiviral NF-κB luciferase reporter construct or p53-luciferase reporter construct (SA Biosciences), and a constitutively expressed renilla luciferase construct (SA Biosciences).

    Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing, Irradiation, Transduction, Luciferase, Construct, Activity Assay