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pbluescript ii ks p21 promoter luc plasmid  (Addgene inc)


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    Addgene inc pbluescript ii ks p21 promoter luc plasmid
    LJ3402 prevents hepatic senescence and mitochondrial dysfunction in aged mice. (A) Representative images of hematoxylin and eosin (H&E) (top), SA‐β‐gal (middle), and Oil Red O (bottom) staining on frozen liver sections from the indicated group. (B) The mRNA levels of marker genes involved in senescence ( p16 , <t>p21</t> , p53 ) and protein levels (γH2AX, p21, p53) in the liver of young and aged mice were determined by RT‐qPCR or immunoblotting ( n = 3; three randomly selected samples per group). (C) Expression of marker genes involved in lipid metabolism ( Srebp1c , Pparα , Cpt1 ) and mitochondrial homeostasis ( Sirt1 , Pgc‐1α , NRF1 , TFAM ) in the liver was determined using RT‐qPCR. (D) Hepatic triglyceride (TG), total cholesterol (TC), and free fatty acid (FFA), and (E) plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ( n = 5; three randomly selected samples per group). (F) Livers separated from mice were incubated in culture medium, and the media were collected after 12 h and 24 h of incubation; the concentrations of TNF‐α and IL‐1β in the media were measured ( n = 5; three randomly selected samples per group). All data are presented as the mean ± SEM. * p < 0.05 and ** p < 0.01. Different lowercase letters above the bars indicate statistically significant differences ( p < 0.05).
    Pbluescript Ii Ks P21 Promoter Luc Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pbluescript/p21+promoter+luciferase+(1002)+(Plasmid+%2349494)/pmc12679060-59-1-9
    Average 93 stars, based on 24 article reviews
    pbluescript ii ks p21 promoter luc plasmid - by Bioz Stars, 2026-11
    93/100 stars

    Images

    1) Product Images from "Lactobacillus johnsonii JNU3402 Ameliorates Age‐Related Liver Dysfunction Through Stimulating PGC ‐1α‐Mediated SIRT1 Expression"

    Article Title: Lactobacillus johnsonii JNU3402 Ameliorates Age‐Related Liver Dysfunction Through Stimulating PGC ‐1α‐Mediated SIRT1 Expression

    Journal: Biofactors (Oxford, England)

    doi: 10.1002/biof.70069

    LJ3402 prevents hepatic senescence and mitochondrial dysfunction in aged mice. (A) Representative images of hematoxylin and eosin (H&E) (top), SA‐β‐gal (middle), and Oil Red O (bottom) staining on frozen liver sections from the indicated group. (B) The mRNA levels of marker genes involved in senescence ( p16 , p21 , p53 ) and protein levels (γH2AX, p21, p53) in the liver of young and aged mice were determined by RT‐qPCR or immunoblotting ( n = 3; three randomly selected samples per group). (C) Expression of marker genes involved in lipid metabolism ( Srebp1c , Pparα , Cpt1 ) and mitochondrial homeostasis ( Sirt1 , Pgc‐1α , NRF1 , TFAM ) in the liver was determined using RT‐qPCR. (D) Hepatic triglyceride (TG), total cholesterol (TC), and free fatty acid (FFA), and (E) plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ( n = 5; three randomly selected samples per group). (F) Livers separated from mice were incubated in culture medium, and the media were collected after 12 h and 24 h of incubation; the concentrations of TNF‐α and IL‐1β in the media were measured ( n = 5; three randomly selected samples per group). All data are presented as the mean ± SEM. * p < 0.05 and ** p < 0.01. Different lowercase letters above the bars indicate statistically significant differences ( p < 0.05).
    Figure Legend Snippet: LJ3402 prevents hepatic senescence and mitochondrial dysfunction in aged mice. (A) Representative images of hematoxylin and eosin (H&E) (top), SA‐β‐gal (middle), and Oil Red O (bottom) staining on frozen liver sections from the indicated group. (B) The mRNA levels of marker genes involved in senescence ( p16 , p21 , p53 ) and protein levels (γH2AX, p21, p53) in the liver of young and aged mice were determined by RT‐qPCR or immunoblotting ( n = 3; three randomly selected samples per group). (C) Expression of marker genes involved in lipid metabolism ( Srebp1c , Pparα , Cpt1 ) and mitochondrial homeostasis ( Sirt1 , Pgc‐1α , NRF1 , TFAM ) in the liver was determined using RT‐qPCR. (D) Hepatic triglyceride (TG), total cholesterol (TC), and free fatty acid (FFA), and (E) plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ( n = 5; three randomly selected samples per group). (F) Livers separated from mice were incubated in culture medium, and the media were collected after 12 h and 24 h of incubation; the concentrations of TNF‐α and IL‐1β in the media were measured ( n = 5; three randomly selected samples per group). All data are presented as the mean ± SEM. * p < 0.05 and ** p < 0.01. Different lowercase letters above the bars indicate statistically significant differences ( p < 0.05).

    Techniques Used: Staining, Marker, Quantitative RT-PCR, Western Blot, Expressing, Clinical Proteomics, Incubation

    SIRT1 modulates LJ3402‐mediated suppression of p53 function in senescent AML12 hepatocytes. (A) Acetylation of p53 after 5 days of treatment with LJ3402‐CM (LJ‐CM) and 50 μM sirtinol during H 2 O 2 ‐induced senescence. (A, C–I) Senescence of AML12 hepatocytes was induced by 1 mM or 750 μM H 2 O 2 for 1 h per day for 7 days. LJ‐CM, 50 μM sirtinol, or 20 μM PFTα were added on days 3–7 during H 2 O 2 exposure. (B) Luciferase reporter activity in HEK293T cells transfected with a reporter plasmid ( p21 promoter–luc) and a p53 expression plasmid. Twelve hours after transfection, cells were treated with LJ‐CM and 50 μM sirtinol for 24 h with or without 100 μM H 2 O 2 . (C) The mRNA and protein levels of p21, p53, PGC‐1α, and SIRT1 measured by RT‐qPCR and immunoblotting, respectively. (D) SA‐β‐gal staining. (E) mRNA levels of genes involved in mitochondrial biogenesis ( NRF1 and TFAM ) and oxidant defense ( CAT and SOD ). (F) ROS levels. (G) Oxygen consumption rates (OCRs) measured at baseline and after treatment with 2.5 μM oligomycin, 1 μM FCCP, or 1 μM rotenone/antimycin A. (H) Oil Red O staining. (I) Intracellular TG levels in senescent AML12 hepatocytes treated with LJ‐CM, sirtinol, and PFTα. All data are presented as the mean ± SEM. Different lowercase letters above the bars indicate statistically significant differences ( p < 0.05).
    Figure Legend Snippet: SIRT1 modulates LJ3402‐mediated suppression of p53 function in senescent AML12 hepatocytes. (A) Acetylation of p53 after 5 days of treatment with LJ3402‐CM (LJ‐CM) and 50 μM sirtinol during H 2 O 2 ‐induced senescence. (A, C–I) Senescence of AML12 hepatocytes was induced by 1 mM or 750 μM H 2 O 2 for 1 h per day for 7 days. LJ‐CM, 50 μM sirtinol, or 20 μM PFTα were added on days 3–7 during H 2 O 2 exposure. (B) Luciferase reporter activity in HEK293T cells transfected with a reporter plasmid ( p21 promoter–luc) and a p53 expression plasmid. Twelve hours after transfection, cells were treated with LJ‐CM and 50 μM sirtinol for 24 h with or without 100 μM H 2 O 2 . (C) The mRNA and protein levels of p21, p53, PGC‐1α, and SIRT1 measured by RT‐qPCR and immunoblotting, respectively. (D) SA‐β‐gal staining. (E) mRNA levels of genes involved in mitochondrial biogenesis ( NRF1 and TFAM ) and oxidant defense ( CAT and SOD ). (F) ROS levels. (G) Oxygen consumption rates (OCRs) measured at baseline and after treatment with 2.5 μM oligomycin, 1 μM FCCP, or 1 μM rotenone/antimycin A. (H) Oil Red O staining. (I) Intracellular TG levels in senescent AML12 hepatocytes treated with LJ‐CM, sirtinol, and PFTα. All data are presented as the mean ± SEM. Different lowercase letters above the bars indicate statistically significant differences ( p < 0.05).

    Techniques Used: Luciferase, Activity Assay, Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, Western Blot, Staining

    Related Articles

    other:

    Article Title: Non-Clinical In Vitro Evaluation of Antibiotic Resistance Gene-Free Plasmids Encoding Human or Murine IL-12 Intended for First-in-Human Clinical Study
    Article Snippet: The murine and human IL-12 fusion genes (p40 and p35 subunits) originated from the commercially available plasmids: human—from the pORF-hIL-12 G2 plasmid, murine—from pORF-mIL-12 (p40:p35) (both: InvivoGen, Toulouse, France).

    Luciferase:

    Article Title: Mechanism of apoptosis activation by Curcumin rescued mutant p53Y220C in human pancreatic cancer.
    Article Snippet: .. Luciferase reporter assay The pGL-3 plasmids containing full-length Mdm2 and p21 promoters purchased from AddGene, MA USA were transfected into the BxPC-3 cells using the Lipofectamine 2000 (Invitrogen, Carlsbad CA, USA). .. After 32 h of transfection, the cells were treated with 3 and 8 μM of curcumin for 16 h, and then the Luciferase activity was detected using the Dual-Luciferase® Reporter Assay System (Promega, WI, USA) following the manufacturer's protocol.

    Reporter Assay:

    Article Title: Mechanism of apoptosis activation by Curcumin rescued mutant p53Y220C in human pancreatic cancer.
    Article Snippet: .. Luciferase reporter assay The pGL-3 plasmids containing full-length Mdm2 and p21 promoters purchased from AddGene, MA USA were transfected into the BxPC-3 cells using the Lipofectamine 2000 (Invitrogen, Carlsbad CA, USA). .. After 32 h of transfection, the cells were treated with 3 and 8 μM of curcumin for 16 h, and then the Luciferase activity was detected using the Dual-Luciferase® Reporter Assay System (Promega, WI, USA) following the manufacturer's protocol.

    Transfection:

    Article Title: Mechanism of apoptosis activation by Curcumin rescued mutant p53Y220C in human pancreatic cancer.
    Article Snippet: .. Luciferase reporter assay The pGL-3 plasmids containing full-length Mdm2 and p21 promoters purchased from AddGene, MA USA were transfected into the BxPC-3 cells using the Lipofectamine 2000 (Invitrogen, Carlsbad CA, USA). .. After 32 h of transfection, the cells were treated with 3 and 8 μM of curcumin for 16 h, and then the Luciferase activity was detected using the Dual-Luciferase® Reporter Assay System (Promega, WI, USA) following the manufacturer's protocol.

    Article Title: Lactobacillus amylovorus KU4 inhibits adipocyte senescence in aged mice through necdin regulation of p53 activity
    Article Snippet: .. Plasmids, pBluescript II KS (+)–p21 promoter Luc (p21-promoter–Luc), were purchased from Addgene (Watertown, MA, USA). pCDNA3–p53 and pCDNA3–p300 were transfected into HEK293T cells, 3T3-L1 adipocytes or primary adipocytes. pCDNA3–NDN was constructed for a previous study. .. Plasmids, pBluescript II KS (+)–p21 promoter Luc (p21-promoter–Luc), were purchased from Addgene (Watertown, MA, USA).

    Article Title: Lactobacillus amylovorus KU4 inhibits adipocyte senescence in aged mice through necdin regulation of p53 activity.
    Article Snippet: .. Plasmids, pBluescript II KS (+)–p21 promoter Luc (p21-promoter–Luc), were purchased from Addgene (Watertown, MA, USA). pCDNA3–p53 and pCDNA3– p300 were transfected into HEK293T cells, 3T3-L1 adipocytes or primary adipocytes. pCDNA3–NDN was constructed for a previous study. ..

    Construct:

    Article Title: Lactobacillus amylovorus KU4 inhibits adipocyte senescence in aged mice through necdin regulation of p53 activity
    Article Snippet: .. Plasmids, pBluescript II KS (+)–p21 promoter Luc (p21-promoter–Luc), were purchased from Addgene (Watertown, MA, USA). pCDNA3–p53 and pCDNA3–p300 were transfected into HEK293T cells, 3T3-L1 adipocytes or primary adipocytes. pCDNA3–NDN was constructed for a previous study. .. Plasmids, pBluescript II KS (+)–p21 promoter Luc (p21-promoter–Luc), were purchased from Addgene (Watertown, MA, USA).

    Article Title: Lactobacillus amylovorus KU4 inhibits adipocyte senescence in aged mice through necdin regulation of p53 activity.
    Article Snippet: .. Plasmids, pBluescript II KS (+)–p21 promoter Luc (p21-promoter–Luc), were purchased from Addgene (Watertown, MA, USA). pCDNA3–p53 and pCDNA3– p300 were transfected into HEK293T cells, 3T3-L1 adipocytes or primary adipocytes. pCDNA3–NDN was constructed for a previous study. ..

    Plasmid Preparation:

    Article Title: Lactobacillus johnsonii JNU3402 Ameliorates Age‐Related Liver Dysfunction Through Stimulating PGC ‐1α‐Mediated SIRT1 Expression
    Article Snippet: .. The pBluescript II KS(+)‐p21 promoter‐luc plasmid was purchased from Addgene (MA, USA) and designated as p21 promoter‐luc. ..

    Article Title: USP39 mediates p21-dependent proliferation and neoplasia of colon cancer cells by regulating the p53/p21/CDC2/cyclin B1 axis.
    Article Snippet: Funding information National Natural Science Foundation of China, Grant/Award Number: 81872045; Public Welfare Research Institutes of Fujian Province, Grant/Award Number: 2020R11010032‐3 Abstract Ubiquitin‐specific protease 39 (USP39) is frequently overexpressed in a variety of cancers, and involved in the regulation of various biological processes, such as cell proliferation, cell cycle progression, apoptosis and pre‐messenger RNA splicing.. Nevertheless, the biological roles and mechanisms of USP39 in colon cancer remain largely unknown.. In this study, we analyzed whether USP39 can be a molecular target for the treatment of colon cancer.

    Article Title: The RNA-binding protein hnRNP E1 regulates p53 and p21 translation via KH1 and KH2 domain interactions with 3' UTR C-rich motifs.
    Article Snippet: For the p53 promoter assay, plasmid pGL2-356bp was used, which was a gift from Wafik El-Deiry (Addgene plasmid # 16292; http://n2t.net/addgene:16292; RRID: Addgene_16292). .. For the p21 promoter assay, pGL2-p21 promoter-Luc was used which was a gift from Martin Walsh (Addgene plasmid # 33021; http://n2t.net/addgene: 33021; RRID: Addgene_33021). .. For p53 5’-UTR-luciferase-3’-UTR expression cassette, 145-pGL3ctrl-3' plasmid was used which was a gift from Michael Kastan (Addgene plasmid # 28175; http://n2t.net/addgene: 28175; RRID: Addgene_28175).

    Article Title: The RNA-binding protein hnRNP E1 regulates p53 and p21 translation via KH1 and KH2 domain interactions with 3′ UTR C-rich motifs
    Article Snippet: For the p53 promoter assay, plasmid pGL2-356 bp was used, which was a gift from Wafik El-Deiry (Addgene plasmid # 16292; http://n2t.net/addgene:16292 ; RRID: Addgene_16292). .. For the p21 promoter assay, pGL2-p21 promoter-Luc was used which was a gift from Martin Walsh (Addgene plasmid # 33021; http://n2t.net/addgene : 33021; RRID: Addgene_33021). .. For p53 5′-UTR-luciferase-3′-UTR expression cassette, 145-pGL3ctrl-3′ plasmid was used which was a gift from Michael Kastan (Addgene plasmid # 28175; http://n2t.net/addgene : 28175; RRID: Addgene_28175).

    Virus:

    Article Title: USP39 mediates p21-dependent proliferation and neoplasia of colon cancer cells by regulating the p53/p21/CDC2/cyclin B1 axis.
    Article Snippet: Funding information National Natural Science Foundation of China, Grant/Award Number: 81872045; Public Welfare Research Institutes of Fujian Province, Grant/Award Number: 2020R11010032‐3 Abstract Ubiquitin‐specific protease 39 (USP39) is frequently overexpressed in a variety of cancers, and involved in the regulation of various biological processes, such as cell proliferation, cell cycle progression, apoptosis and pre‐messenger RNA splicing.. Nevertheless, the biological roles and mechanisms of USP39 in colon cancer remain largely unknown.. In this study, we analyzed whether USP39 can be a molecular target for the treatment of colon cancer.

    Promoter Assay:

    Article Title: The RNA-binding protein hnRNP E1 regulates p53 and p21 translation via KH1 and KH2 domain interactions with 3' UTR C-rich motifs.
    Article Snippet: For the p53 promoter assay, plasmid pGL2-356bp was used, which was a gift from Wafik El-Deiry (Addgene plasmid # 16292; http://n2t.net/addgene:16292; RRID: Addgene_16292). .. For the p21 promoter assay, pGL2-p21 promoter-Luc was used which was a gift from Martin Walsh (Addgene plasmid # 33021; http://n2t.net/addgene: 33021; RRID: Addgene_33021). .. For p53 5’-UTR-luciferase-3’-UTR expression cassette, 145-pGL3ctrl-3' plasmid was used which was a gift from Michael Kastan (Addgene plasmid # 28175; http://n2t.net/addgene: 28175; RRID: Addgene_28175).

    Article Title: The RNA-binding protein hnRNP E1 regulates p53 and p21 translation via KH1 and KH2 domain interactions with 3′ UTR C-rich motifs
    Article Snippet: For the p53 promoter assay, plasmid pGL2-356 bp was used, which was a gift from Wafik El-Deiry (Addgene plasmid # 16292; http://n2t.net/addgene:16292 ; RRID: Addgene_16292). .. For the p21 promoter assay, pGL2-p21 promoter-Luc was used which was a gift from Martin Walsh (Addgene plasmid # 33021; http://n2t.net/addgene : 33021; RRID: Addgene_33021). .. For p53 5′-UTR-luciferase-3′-UTR expression cassette, 145-pGL3ctrl-3′ plasmid was used which was a gift from Michael Kastan (Addgene plasmid # 28175; http://n2t.net/addgene : 28175; RRID: Addgene_28175).



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    Pbluescript Ii Ks P21 Promoter Luc Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    LJ3402 prevents hepatic senescence and mitochondrial dysfunction in aged mice. (A) Representative images of hematoxylin and eosin (H&E) (top), SA‐β‐gal (middle), and Oil Red O (bottom) staining on frozen liver sections from the indicated group. (B) The mRNA levels of marker genes involved in senescence ( p16 , p21 , p53 ) and protein levels (γH2AX, p21, p53) in the liver of young and aged mice were determined by RT‐qPCR or immunoblotting ( n = 3; three randomly selected samples per group). (C) Expression of marker genes involved in lipid metabolism ( Srebp1c , Pparα , Cpt1 ) and mitochondrial homeostasis ( Sirt1 , Pgc‐1α , NRF1 , TFAM ) in the liver was determined using RT‐qPCR. (D) Hepatic triglyceride (TG), total cholesterol (TC), and free fatty acid (FFA), and (E) plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ( n = 5; three randomly selected samples per group). (F) Livers separated from mice were incubated in culture medium, and the media were collected after 12 h and 24 h of incubation; the concentrations of TNF‐α and IL‐1β in the media were measured ( n = 5; three randomly selected samples per group). All data are presented as the mean ± SEM. * p < 0.05 and ** p < 0.01. Different lowercase letters above the bars indicate statistically significant differences ( p < 0.05).

    Journal: Biofactors (Oxford, England)

    Article Title: Lactobacillus johnsonii JNU3402 Ameliorates Age‐Related Liver Dysfunction Through Stimulating PGC ‐1α‐Mediated SIRT1 Expression

    doi: 10.1002/biof.70069

    Figure Lengend Snippet: LJ3402 prevents hepatic senescence and mitochondrial dysfunction in aged mice. (A) Representative images of hematoxylin and eosin (H&E) (top), SA‐β‐gal (middle), and Oil Red O (bottom) staining on frozen liver sections from the indicated group. (B) The mRNA levels of marker genes involved in senescence ( p16 , p21 , p53 ) and protein levels (γH2AX, p21, p53) in the liver of young and aged mice were determined by RT‐qPCR or immunoblotting ( n = 3; three randomly selected samples per group). (C) Expression of marker genes involved in lipid metabolism ( Srebp1c , Pparα , Cpt1 ) and mitochondrial homeostasis ( Sirt1 , Pgc‐1α , NRF1 , TFAM ) in the liver was determined using RT‐qPCR. (D) Hepatic triglyceride (TG), total cholesterol (TC), and free fatty acid (FFA), and (E) plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ( n = 5; three randomly selected samples per group). (F) Livers separated from mice were incubated in culture medium, and the media were collected after 12 h and 24 h of incubation; the concentrations of TNF‐α and IL‐1β in the media were measured ( n = 5; three randomly selected samples per group). All data are presented as the mean ± SEM. * p < 0.05 and ** p < 0.01. Different lowercase letters above the bars indicate statistically significant differences ( p < 0.05).

    Article Snippet: The pBluescript II KS(+)‐p21 promoter‐luc plasmid was purchased from Addgene (MA, USA) and designated as p21 promoter‐luc.

    Techniques: Staining, Marker, Quantitative RT-PCR, Western Blot, Expressing, Clinical Proteomics, Incubation

    SIRT1 modulates LJ3402‐mediated suppression of p53 function in senescent AML12 hepatocytes. (A) Acetylation of p53 after 5 days of treatment with LJ3402‐CM (LJ‐CM) and 50 μM sirtinol during H 2 O 2 ‐induced senescence. (A, C–I) Senescence of AML12 hepatocytes was induced by 1 mM or 750 μM H 2 O 2 for 1 h per day for 7 days. LJ‐CM, 50 μM sirtinol, or 20 μM PFTα were added on days 3–7 during H 2 O 2 exposure. (B) Luciferase reporter activity in HEK293T cells transfected with a reporter plasmid ( p21 promoter–luc) and a p53 expression plasmid. Twelve hours after transfection, cells were treated with LJ‐CM and 50 μM sirtinol for 24 h with or without 100 μM H 2 O 2 . (C) The mRNA and protein levels of p21, p53, PGC‐1α, and SIRT1 measured by RT‐qPCR and immunoblotting, respectively. (D) SA‐β‐gal staining. (E) mRNA levels of genes involved in mitochondrial biogenesis ( NRF1 and TFAM ) and oxidant defense ( CAT and SOD ). (F) ROS levels. (G) Oxygen consumption rates (OCRs) measured at baseline and after treatment with 2.5 μM oligomycin, 1 μM FCCP, or 1 μM rotenone/antimycin A. (H) Oil Red O staining. (I) Intracellular TG levels in senescent AML12 hepatocytes treated with LJ‐CM, sirtinol, and PFTα. All data are presented as the mean ± SEM. Different lowercase letters above the bars indicate statistically significant differences ( p < 0.05).

    Journal: Biofactors (Oxford, England)

    Article Title: Lactobacillus johnsonii JNU3402 Ameliorates Age‐Related Liver Dysfunction Through Stimulating PGC ‐1α‐Mediated SIRT1 Expression

    doi: 10.1002/biof.70069

    Figure Lengend Snippet: SIRT1 modulates LJ3402‐mediated suppression of p53 function in senescent AML12 hepatocytes. (A) Acetylation of p53 after 5 days of treatment with LJ3402‐CM (LJ‐CM) and 50 μM sirtinol during H 2 O 2 ‐induced senescence. (A, C–I) Senescence of AML12 hepatocytes was induced by 1 mM or 750 μM H 2 O 2 for 1 h per day for 7 days. LJ‐CM, 50 μM sirtinol, or 20 μM PFTα were added on days 3–7 during H 2 O 2 exposure. (B) Luciferase reporter activity in HEK293T cells transfected with a reporter plasmid ( p21 promoter–luc) and a p53 expression plasmid. Twelve hours after transfection, cells were treated with LJ‐CM and 50 μM sirtinol for 24 h with or without 100 μM H 2 O 2 . (C) The mRNA and protein levels of p21, p53, PGC‐1α, and SIRT1 measured by RT‐qPCR and immunoblotting, respectively. (D) SA‐β‐gal staining. (E) mRNA levels of genes involved in mitochondrial biogenesis ( NRF1 and TFAM ) and oxidant defense ( CAT and SOD ). (F) ROS levels. (G) Oxygen consumption rates (OCRs) measured at baseline and after treatment with 2.5 μM oligomycin, 1 μM FCCP, or 1 μM rotenone/antimycin A. (H) Oil Red O staining. (I) Intracellular TG levels in senescent AML12 hepatocytes treated with LJ‐CM, sirtinol, and PFTα. All data are presented as the mean ± SEM. Different lowercase letters above the bars indicate statistically significant differences ( p < 0.05).

    Article Snippet: The pBluescript II KS(+)‐p21 promoter‐luc plasmid was purchased from Addgene (MA, USA) and designated as p21 promoter‐luc.

    Techniques: Luciferase, Activity Assay, Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, Western Blot, Staining