human pgc-1α Search Results


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  • 91
    Addgene inc human pgc 1α
    Overexpression of <t>pgc-1α</t> suppresses 4-ClBQ induced down-regulation of catalase expression in HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure PGC-1α (A) mRNA and (B) protein levels in vector-alone and PGC-1α
    Human Pgc 1α, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Horizon Discovery human pgc 1α
    Proposed model for the protective effects of adiponectin on dystrophic muscle. Signal transduction mediating ApN protection on dystrophic muscle: binding of ApN to AdipoR1 activates the <t>AMPK/SIRT1/PGC-1α</t> pathway. Briefly, ApN leads to AMPK phosphorylation/activation. P-AMPK in turn phosphorylates PGC-1α and indirectly increases the expression of SIRT1 (through rising NAD+/NADH ratio). SIRT1 in turn deacetylates and fully activates PGC-1α. Next, PGC-1α represses NF-κB activity by dephosphorylation of the p65 subunit [ 38 ], while SIRT1 represses it by deacetylation [ 39 ]. This results in decreased muscle inflammation/oxidative stress and improved myogenic program as well as enhanced utrophin expression and oxidative capacity, both processes helping rescue the dystrophic phenotype. Green arrow , stimulation; red arrow , inhibition
    Human Pgc 1α, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 88/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam rabbit anti human peroxisome proliferator activated receptor gamma coactivator 1 alpha pgc1α antibody
    Resveratrol (RSV) rescued the cell aggregate formation and osteogenic differentiation of N-PDLSCs under inflammatory cytokine tumor necrosis factor <t>alpha</t> (TNF-α) treatment. a – c Histological analysis of PDLSC aggregates ( a ) showed that N-PDLSCs under TNF-α treatment formed thinner aggregates ( b ) with less collagen deposition ( c ), which could be rescued by RSV application. Bars: 50 μm. d Osteogenesis of PDLSC aggregates demonstrated by ALP (top) and alizarin red staining (bottom). Bars: 5 mm (top) and 50 μm (bottom). e , f RSV treatment restored the ALP activity ( e ) and mineralizing activity ( f ) of N-PDLSC aggregates under TNF-α application. g Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of the osteogenic marker genes Alp , Runt-related transcription factor 2 ( Runx2 ), Osteocalcin ( Ocn ), and Collagen I ( Col1 ) in PDLSC aggregates after osteogenic induction. The data showed that RSV rescued the osteogenic potential of N-PDLSC aggregates under TNF-α treatment. h Western blot analysis of the extracellular matrix (ECM) and osteogenic marker genes Col1, Periostin, and Runx2 in PDLSC aggregates. i Western blot analysis of molecular targets related to key signaling pathways in PDLSC aggregates: the Sirtuin 1 (Sirt1)-peroxisome <t>proliferator-activated</t> receptor <t>gamma</t> <t>coactivator-1</t> alpha <t>(Pgc1α)</t> pathway; the nuclear factor kappaB (NFκB) pathway; and the adenosine monophosphate-activated protein kinase (AMPK) pathway. j Western blot analysis showed that RSV improved osteogenesis and ECM deposition in N-PDLSC aggregates under TNF-α treatment, possibly by inhibiting the NFκB pathway and stimulating the Pgc1α and AMPK pathways. TNF-α was applied at 10 ng/ml, and RSV was applied at 10 nM in all the experiments. n = 6 per group ( a – f ) and n = 3 per group ( g – j ). The data represent the mean ± SD. * P
    Rabbit Anti Human Peroxisome Proliferator Activated Receptor Gamma Coactivator 1 Alpha Pgc1α Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology antibody against human pgc 1α
    Effects of <t>PGC-1α</t> knockdown on the mitochondrial membrane potential in the HEC-1A cells. Notes: ( A ) Cells were stained with JC-1 and analyzed with flow cytometry after 72 hours of transfection with siRNA-2 or NC siRNA. siRNA-2 treatment increased the number of cells with low Δ ψ m . ( B ) Quantification of the results showed that siRNA-2 treatment increased the number of cells with low Δ ψ m . Results are shown as the mean ± SD values of three independent experiments. * P
    Antibody Against Human Pgc 1α, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Cell Signaling Technology Inc anti human pgc 1α antibody
    Immunofluorescence staining of apoptotic nuclei ( green ) and mitochondria ( red ) in human musculoskeletal cell lines after <t>PGC-1α</t> plasmid and TFAM siRNA co-transfection. (a) Nara-H, a human MFH cell line and (b) KTHOS, a human osteosarcoma cell line.
    Anti Human Pgc 1α Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti human pgc 1α polyclonal antibody
    Palmitic acid induced type 2 diabetes-like features in HepG2 cells. (A) Lipid level was increased in PA concentration-dependent manner in PA-treated cells determined by Nile Red staining. (B) Lipid metabolism-related proteins, PPARα and <t>PGC-1α,</t> were reduced in PA-treated cells. (C) Transcription levels of PPARα and PGC-1α were decreased by PA treatment. (D) Western blot results show that preproinsulin levels increased, and IRS2 and GLUT2 expression decreased in PA-treated cells in a concentration-dependent manner. (E) Transcription levels of insulin elevated and of IRS2 and GLUT2 declined following PA treatment. The upper panel shows representative electrophorogram of the gene bands from RT-PCR and lower pane ls show the corresponding histograms from real-time qPCR in C and E. *p
    Anti Human Pgc 1α Polyclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Abcam anti human pgc 1α
    Palmitic acid induced type 2 diabetes-like features in HepG2 cells. (A) Lipid level was increased in PA concentration-dependent manner in PA-treated cells determined by Nile Red staining. (B) Lipid metabolism-related proteins, PPARα and <t>PGC-1α,</t> were reduced in PA-treated cells. (C) Transcription levels of PPARα and PGC-1α were decreased by PA treatment. (D) Western blot results show that preproinsulin levels increased, and IRS2 and GLUT2 expression decreased in PA-treated cells in a concentration-dependent manner. (E) Transcription levels of insulin elevated and of IRS2 and GLUT2 declined following PA treatment. The upper panel shows representative electrophorogram of the gene bands from RT-PCR and lower pane ls show the corresponding histograms from real-time qPCR in C and E. *p
    Anti Human Pgc 1α, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Biorbyt anti pgc 1α rabbit anti human polyclonal
    Graphical summary of inter-variable connections. Figure 2 illustrates statistical connections of variables of interest. Bivariate correlations were calculated for the entire study population using the model of Spearman’s rho. R coefficients and p-values (p ≤ 0.05; in brackets) are presented in the grey box and summarised in a graphical model. For ease of reading, lifestyle factors have not been incorporated in detail, and are therefore only abstracted (bottom of figure). Abbreviations: UCB: unconjugated bilirubin; UGT1A1 : UGT1A1 genotype; pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; <t>PgC</t> <t>1α:</t> Peroxisome proliferator-activated receptor c coactivator 1; Sirt-1: Sirtuin-1; FGF-21: Fibroblast growth factor 21; T3: Free triiodothyronine; BMI: Body mass index; LBM: Lean body mass; HbA1c: Glycated haemoglobin A1c; TChol: Total cholesterol; HDL: High density lipoprotein cholesterol; LDL: Low density lipoprotein cholesterol; TG: Triglyceride; LPA2: Lipoprotein A2; ApoA1: Apolipoprotein A1; ApoB: Apolipoprotein B.
    Anti Pgc 1α Rabbit Anti Human Polyclonal, supplied by Biorbyt, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Addgene inc human myc pgc1α plasmid
    Graphical summary of inter-variable connections. Figure 2 illustrates statistical connections of variables of interest. Bivariate correlations were calculated for the entire study population using the model of Spearman’s rho. R coefficients and p-values (p ≤ 0.05; in brackets) are presented in the grey box and summarised in a graphical model. For ease of reading, lifestyle factors have not been incorporated in detail, and are therefore only abstracted (bottom of figure). Abbreviations: UCB: unconjugated bilirubin; UGT1A1 : UGT1A1 genotype; pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; <t>PgC</t> <t>1α:</t> Peroxisome proliferator-activated receptor c coactivator 1; Sirt-1: Sirtuin-1; FGF-21: Fibroblast growth factor 21; T3: Free triiodothyronine; BMI: Body mass index; LBM: Lean body mass; HbA1c: Glycated haemoglobin A1c; TChol: Total cholesterol; HDL: High density lipoprotein cholesterol; LDL: Low density lipoprotein cholesterol; TG: Triglyceride; LPA2: Lipoprotein A2; ApoA1: Apolipoprotein A1; ApoB: Apolipoprotein B.
    Human Myc Pgc1α Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Addgene inc human pgc 1α plasmid dna
    Overexpression of <t>pgc-1α</t> suppresses 4-ClBQ induced down-regulation of catalase expression in HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure PGC-1α (A) mRNA and (B) protein levels in vector-alone and PGC-1α
    Human Pgc 1α Plasmid Dna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    GenePharma Company human pgc 1α mrna
    Effects of <t>PGC-1α</t> knockdown on the mitochondrial membrane potential in the HEC-1A cells. Notes: ( A ) Cells were stained with JC-1 and analyzed with flow cytometry after 72 hours of transfection with siRNA-2 or NC siRNA. siRNA-2 treatment increased the number of cells with low Δ ψ m . ( B ) Quantification of the results showed that siRNA-2 treatment increased the number of cells with low Δ ψ m . Results are shown as the mean ± SD values of three independent experiments. * P
    Human Pgc 1α Mrna, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Addgene inc human pgc 1α pcdna4 plasmid
    Anti-apoptotic effect of <t>PGC-1α.</t> Stable cells were treated with 0.5 mM H 2 O 2 for 6 h. ( A ) The expression bands of apoptotic proteins in Mock and PGC-1α-stable cells were compared via western blotting. Each bar graph represents the expression of PGC-1α ( B ), ratio of phosphorylated p53 at Ser 15 to total p53 ( C ), the level of activated caspase 3 (ratio of cleaved caspase 3 to caspase 3 ( D ), and the level of cytochrome C release from mitochondria to cytosol ( E ). β-actin levels were analyzed as internal controls. GAPDH and complex V α were used as internal controls in cytosol and in mitochondria fraction, respectively. Full-length blots of each tested protein are reported in Supplementary Figure S4 . Error bars denote the mean ± S.D. of triplicate samples. * p
    Human Pgc 1α Pcdna4 Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Addgene inc human pgc 1α promoter luc
    Mutant huntingtin results in repressed transcriptional activities, and reduced PPARγ activity is independent of the protein level. B3 and E4 STHdh Q7 cells and 1A and 6L STHdh Q111 cells were transiently transfected with PPRE, CRE, or <t>PGC-1α</t> promoter luciferase reporter plasmids. The basal activities of PPRE ( A ), CRE ( B ), and PGC-1α promoter ( C ) reporters were dramatically reduced in both 1A and 6L STHdh Q111 cells. n = 3–4 Data shown are mean ± SE. D , PPARγ protein levels were variable in B3 and E4 STHdh Q7 cells and 1A and 6L STHdh Q111 cells, while the original STHdh Q111 cells exhibited lower levels of PPARγ than the original STHdh Q7 cells. Sixty micrograms of protein was run in each lane.
    Human Pgc 1α Promoter Luc, supplied by Addgene inc, used in various techniques. Bioz Stars score: 85/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Vector Biolabs human pgc 1α
    Schematic model of Erk5 protection against metabolic stress. FFA induces escalated ROS production from Gp91phox, which activates caplain-1, leading to a breakdown of Erk5. Loss of Erk5 and resultant <t>Pgc-1α</t> downregulation impose detrimental impacts on mitochondrial functions through accumulation of toxic lipids and vicious cycle of ROS. Prevention of Erk5 degradation or Erk5 restoration by AAV9 approach revives mitochondrial functions
    Human Pgc 1α, supplied by Vector Biolabs, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human phospho pgc1 alpha s571 antibody
    Schematic model of Erk5 protection against metabolic stress. FFA induces escalated ROS production from Gp91phox, which activates caplain-1, leading to a breakdown of Erk5. Loss of Erk5 and resultant <t>Pgc-1α</t> downregulation impose detrimental impacts on mitochondrial functions through accumulation of toxic lipids and vicious cycle of ROS. Prevention of Erk5 degradation or Erk5 restoration by AAV9 approach revives mitochondrial functions
    Human Phospho Pgc1 Alpha S571 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology rabbit anti human pgc 1α polyclonal antibodies
    Identification of LDLR promoter region that is responsible for <t>PGC-1α-mediated</t> suppression. The original pLR1563-luc and the indicated series of 5'-deletion constructs linked to luciferase were cotransfected into HepG2 cells with LacZ expression
    Rabbit Anti Human Pgc 1α Polyclonal Antibodies, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti human pgc 1α polyclonal antibodies/product/Santa Cruz Biotechnology
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    88
    Abcam mouse anti human pgc 1α
    Changes in VEGF protein levels Effect of <t>PGC-1α</t> siRNA on VEGF protein production in hRVECs at 16h of normoxia and hypoxia after transfection were detected by ELISA. The VEGF protein production in siPGC-1α groups were significantly downregulated when compared with control siRNA groups under normoxia and hypoxia. b P
    Mouse Anti Human Pgc 1α, supplied by Abcam, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene mice human pgc 1α cdna
    Endothelial <t>PGC-1α</t> overexpression protects from endothelial dysfunction. ( A ) PGC-1α endothelial specific transgene construct with representative expression of human (h) and mouse (m) PGC-1α mRNA in MLECs from the indicated genotype. ( B ) MLEC mRNA quantification (N = 3, * P
    Mice Human Pgc 1α Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    OriGene kb human pgc 1α hpgc 1α cdna
    Plasmid construct for generation of <t>actin-PGC-1α</t> transgenic mice. A Not I site was inserted at the vector Xba I site and a Sal I site inserted at the vector Hind III site on the pCAGGS plasmid. Human peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC-1α) <t>cDNA</t> was inserted in the Not I site and Sal I digestion released the fragment containing the cytomegalovirus immediate-early (CMV-IE) enhancer, chicken β-actin promoter, and <t>hPGC-1α</t> that was used for microinjection.
    Kb Human Pgc 1α Hpgc 1α Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Overexpression of pgc-1α suppresses 4-ClBQ induced down-regulation of catalase expression in HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure PGC-1α (A) mRNA and (B) protein levels in vector-alone and PGC-1α

    Journal: Toxicology in vitro : an international journal published in association with BIBRA

    Article Title: Down-regulation of peroxisome proliferator activated receptor γ coactivator 1α induces oxidative stress and toxicity of 1-(4-Chlorophenyl)-benzo-2,5-quinone in HaCaT human keratinocytes

    doi: 10.1016/j.tiv.2015.05.009

    Figure Lengend Snippet: Overexpression of pgc-1α suppresses 4-ClBQ induced down-regulation of catalase expression in HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure PGC-1α (A) mRNA and (B) protein levels in vector-alone and PGC-1α

    Article Snippet: Human pgc-1α was transiently overexpressed in HaCaT cells using human PGC-1α plasmid DNA (Addgene, plasmid No. 10974; Cambridge, MA).

    Techniques: Over Expression, Pyrolysis Gas Chromatography, Expressing, Quantitative RT-PCR, Plasmid Preparation

    pgc-1α expression is inhibited in 4-ClBQ treated HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure pgc-1α (A) mRNA and (B) protein levels in control and 4-ClBQ treated cells at 24 h after the addition of 4-ClBQ.

    Journal: Toxicology in vitro : an international journal published in association with BIBRA

    Article Title: Down-regulation of peroxisome proliferator activated receptor γ coactivator 1α induces oxidative stress and toxicity of 1-(4-Chlorophenyl)-benzo-2,5-quinone in HaCaT human keratinocytes

    doi: 10.1016/j.tiv.2015.05.009

    Figure Lengend Snippet: pgc-1α expression is inhibited in 4-ClBQ treated HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure pgc-1α (A) mRNA and (B) protein levels in control and 4-ClBQ treated cells at 24 h after the addition of 4-ClBQ.

    Article Snippet: Human pgc-1α was transiently overexpressed in HaCaT cells using human PGC-1α plasmid DNA (Addgene, plasmid No. 10974; Cambridge, MA).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Quantitative RT-PCR

    Overexpression of pgc-1α suppresses 4-ClBQ induced oxidative stress and toxicity in HaCaT cells. Flow cytometry measurements of MitoSOX Red oxidation were used to determine cellular ROS levels in control and 4-ClBQ treated vector-alone and pgc-1α

    Journal: Toxicology in vitro : an international journal published in association with BIBRA

    Article Title: Down-regulation of peroxisome proliferator activated receptor γ coactivator 1α induces oxidative stress and toxicity of 1-(4-Chlorophenyl)-benzo-2,5-quinone in HaCaT human keratinocytes

    doi: 10.1016/j.tiv.2015.05.009

    Figure Lengend Snippet: Overexpression of pgc-1α suppresses 4-ClBQ induced oxidative stress and toxicity in HaCaT cells. Flow cytometry measurements of MitoSOX Red oxidation were used to determine cellular ROS levels in control and 4-ClBQ treated vector-alone and pgc-1α

    Article Snippet: Human pgc-1α was transiently overexpressed in HaCaT cells using human PGC-1α plasmid DNA (Addgene, plasmid No. 10974; Cambridge, MA).

    Techniques: Over Expression, Pyrolysis Gas Chromatography, Flow Cytometry, Cytometry, Plasmid Preparation

    Proposed model for the protective effects of adiponectin on dystrophic muscle. Signal transduction mediating ApN protection on dystrophic muscle: binding of ApN to AdipoR1 activates the AMPK/SIRT1/PGC-1α pathway. Briefly, ApN leads to AMPK phosphorylation/activation. P-AMPK in turn phosphorylates PGC-1α and indirectly increases the expression of SIRT1 (through rising NAD+/NADH ratio). SIRT1 in turn deacetylates and fully activates PGC-1α. Next, PGC-1α represses NF-κB activity by dephosphorylation of the p65 subunit [ 38 ], while SIRT1 represses it by deacetylation [ 39 ]. This results in decreased muscle inflammation/oxidative stress and improved myogenic program as well as enhanced utrophin expression and oxidative capacity, both processes helping rescue the dystrophic phenotype. Green arrow , stimulation; red arrow , inhibition

    Journal: Skeletal Muscle

    Article Title: Involvement of adiponectin in the pathogenesis of dystrophinopathy

    doi: 10.1186/s13395-015-0051-9

    Figure Lengend Snippet: Proposed model for the protective effects of adiponectin on dystrophic muscle. Signal transduction mediating ApN protection on dystrophic muscle: binding of ApN to AdipoR1 activates the AMPK/SIRT1/PGC-1α pathway. Briefly, ApN leads to AMPK phosphorylation/activation. P-AMPK in turn phosphorylates PGC-1α and indirectly increases the expression of SIRT1 (through rising NAD+/NADH ratio). SIRT1 in turn deacetylates and fully activates PGC-1α. Next, PGC-1α represses NF-κB activity by dephosphorylation of the p65 subunit [ 38 ], while SIRT1 represses it by deacetylation [ 39 ]. This results in decreased muscle inflammation/oxidative stress and improved myogenic program as well as enhanced utrophin expression and oxidative capacity, both processes helping rescue the dystrophic phenotype. Green arrow , stimulation; red arrow , inhibition

    Article Snippet: Briefly, 3.105 cells/well were transfected with either the On-Targetplus non-targeting pool siRNAs (negative control, NT siRNAs), or a specific On-Targetplus siRNA SMARTpool against human AdipoR1 (50 nM) or human SIRT1 (50 nM) or human PGC-1α (70 nM) (all from Dharmacon, Thermo Fisher Scientific) using 7 μl Lipofectamine RNAiMAX reagent (Life Technologies) for 24 h. siRNA silencing was effective, ranging from 70 to 95 % in all experiments.

    Techniques: Transduction, Binding Assay, Pyrolysis Gas Chromatography, Activation Assay, Expressing, Activity Assay, De-Phosphorylation Assay, Inhibition

    Effects of adiponectin on AMPK signaling pathway and NF-κB activity in tibialis anterior muscles of mdx mice. The expression of P-AMPK (phosphorylated form) ( a ) and SIRT1 ( b ) was analyzed by Western blotting in muscles from the three groups of mice. c Densitometry of immunoprecipitation experiments performed on skeletal muscle lysates, using anti-PGC-1α antibody for precipitation and anti-acetyl-lysine antibody for immunoblotting. d mRNA levels of Myh7, a marker of slow twitch, oxidative myofibers. e mRNA levels of Myh1, a marker of fast twitch, glycolytic myofibers. f mRNA and g protein levels of utrophin A (UTRN) with a representative Western blot and Ponceau S stain. h Immunodetection of NF-κB (p65) in tibialis anterior sections; some positive marked nuclei ( brown color) are indicated by arrows . Scale bar = 25 μm. i Quantification of p65 immunolabeling in myofiber nuclei (expressed as percent of total nuclei) in sections (as those shown in g ). j Immunoblotting of phosphorylated NF-κB p65 in the same muscles. Levels of P-AMPK, SIRT1, and Acetyl-Lys were normalized to AMPK, actin, and PGC-1α levels, respectively. mRNA levels were normalized to cyclophilin, utrophin A protein levels to Ponceau, and P-p65 to Actin. The subsequent ratios were presented as relative expression compared to WT values. Results are means ± SD; n = 6 mice per group. * p

    Journal: Skeletal Muscle

    Article Title: Involvement of adiponectin in the pathogenesis of dystrophinopathy

    doi: 10.1186/s13395-015-0051-9

    Figure Lengend Snippet: Effects of adiponectin on AMPK signaling pathway and NF-κB activity in tibialis anterior muscles of mdx mice. The expression of P-AMPK (phosphorylated form) ( a ) and SIRT1 ( b ) was analyzed by Western blotting in muscles from the three groups of mice. c Densitometry of immunoprecipitation experiments performed on skeletal muscle lysates, using anti-PGC-1α antibody for precipitation and anti-acetyl-lysine antibody for immunoblotting. d mRNA levels of Myh7, a marker of slow twitch, oxidative myofibers. e mRNA levels of Myh1, a marker of fast twitch, glycolytic myofibers. f mRNA and g protein levels of utrophin A (UTRN) with a representative Western blot and Ponceau S stain. h Immunodetection of NF-κB (p65) in tibialis anterior sections; some positive marked nuclei ( brown color) are indicated by arrows . Scale bar = 25 μm. i Quantification of p65 immunolabeling in myofiber nuclei (expressed as percent of total nuclei) in sections (as those shown in g ). j Immunoblotting of phosphorylated NF-κB p65 in the same muscles. Levels of P-AMPK, SIRT1, and Acetyl-Lys were normalized to AMPK, actin, and PGC-1α levels, respectively. mRNA levels were normalized to cyclophilin, utrophin A protein levels to Ponceau, and P-p65 to Actin. The subsequent ratios were presented as relative expression compared to WT values. Results are means ± SD; n = 6 mice per group. * p

    Article Snippet: Briefly, 3.105 cells/well were transfected with either the On-Targetplus non-targeting pool siRNAs (negative control, NT siRNAs), or a specific On-Targetplus siRNA SMARTpool against human AdipoR1 (50 nM) or human SIRT1 (50 nM) or human PGC-1α (70 nM) (all from Dharmacon, Thermo Fisher Scientific) using 7 μl Lipofectamine RNAiMAX reagent (Life Technologies) for 24 h. siRNA silencing was effective, ranging from 70 to 95 % in all experiments.

    Techniques: Activity Assay, Mouse Assay, Expressing, Western Blot, Immunoprecipitation, Pyrolysis Gas Chromatography, Marker, Staining, Immunodetection, Immunolabeling

    Effects of adiponectin on inflammatory markers in human skeletal muscle. TNFα ( a ) and IL-6 ( b ) mRNAs in primary culture of human myotubes: cells were treated with or without ApN, while being or not challenged with TNFα and IFNγ. TNFα ( c ) and IκB ( d ) mRNAs in human myotubes, which were transfected with siRNAs against AdipoR1, SIRT1, PGC-1α, or a negative (non-targeting, NT) control. After transfection, cells were treated with or without ApN, while being challenged with TNFα/IFNγ. mRNA levels were normalized to TATA box-binding protein. The subsequent ratios are presented as relative expression compared to control conditions [i.e., no TNFα/IFNγ and no ApN ( a , b ); NT siRNA without ApN ( c , d )]. e Utrophin A mRNAs in human myotubes, which were treated with or without ApN, while being challenged with TNFα and IFNγ. mRNA levels were normalized as above. In this last graph, each patient is represented as being its own control (statistical analysis was performed on raw paired data). Data are means ± SD; n = 5–6 different subjects. ** p

    Journal: Skeletal Muscle

    Article Title: Involvement of adiponectin in the pathogenesis of dystrophinopathy

    doi: 10.1186/s13395-015-0051-9

    Figure Lengend Snippet: Effects of adiponectin on inflammatory markers in human skeletal muscle. TNFα ( a ) and IL-6 ( b ) mRNAs in primary culture of human myotubes: cells were treated with or without ApN, while being or not challenged with TNFα and IFNγ. TNFα ( c ) and IκB ( d ) mRNAs in human myotubes, which were transfected with siRNAs against AdipoR1, SIRT1, PGC-1α, or a negative (non-targeting, NT) control. After transfection, cells were treated with or without ApN, while being challenged with TNFα/IFNγ. mRNA levels were normalized to TATA box-binding protein. The subsequent ratios are presented as relative expression compared to control conditions [i.e., no TNFα/IFNγ and no ApN ( a , b ); NT siRNA without ApN ( c , d )]. e Utrophin A mRNAs in human myotubes, which were treated with or without ApN, while being challenged with TNFα and IFNγ. mRNA levels were normalized as above. In this last graph, each patient is represented as being its own control (statistical analysis was performed on raw paired data). Data are means ± SD; n = 5–6 different subjects. ** p

    Article Snippet: Briefly, 3.105 cells/well were transfected with either the On-Targetplus non-targeting pool siRNAs (negative control, NT siRNAs), or a specific On-Targetplus siRNA SMARTpool against human AdipoR1 (50 nM) or human SIRT1 (50 nM) or human PGC-1α (70 nM) (all from Dharmacon, Thermo Fisher Scientific) using 7 μl Lipofectamine RNAiMAX reagent (Life Technologies) for 24 h. siRNA silencing was effective, ranging from 70 to 95 % in all experiments.

    Techniques: Transfection, Pyrolysis Gas Chromatography, Binding Assay, Expressing

    Resveratrol (RSV) rescued the cell aggregate formation and osteogenic differentiation of N-PDLSCs under inflammatory cytokine tumor necrosis factor alpha (TNF-α) treatment. a – c Histological analysis of PDLSC aggregates ( a ) showed that N-PDLSCs under TNF-α treatment formed thinner aggregates ( b ) with less collagen deposition ( c ), which could be rescued by RSV application. Bars: 50 μm. d Osteogenesis of PDLSC aggregates demonstrated by ALP (top) and alizarin red staining (bottom). Bars: 5 mm (top) and 50 μm (bottom). e , f RSV treatment restored the ALP activity ( e ) and mineralizing activity ( f ) of N-PDLSC aggregates under TNF-α application. g Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of the osteogenic marker genes Alp , Runt-related transcription factor 2 ( Runx2 ), Osteocalcin ( Ocn ), and Collagen I ( Col1 ) in PDLSC aggregates after osteogenic induction. The data showed that RSV rescued the osteogenic potential of N-PDLSC aggregates under TNF-α treatment. h Western blot analysis of the extracellular matrix (ECM) and osteogenic marker genes Col1, Periostin, and Runx2 in PDLSC aggregates. i Western blot analysis of molecular targets related to key signaling pathways in PDLSC aggregates: the Sirtuin 1 (Sirt1)-peroxisome proliferator-activated receptor gamma coactivator-1 alpha (Pgc1α) pathway; the nuclear factor kappaB (NFκB) pathway; and the adenosine monophosphate-activated protein kinase (AMPK) pathway. j Western blot analysis showed that RSV improved osteogenesis and ECM deposition in N-PDLSC aggregates under TNF-α treatment, possibly by inhibiting the NFκB pathway and stimulating the Pgc1α and AMPK pathways. TNF-α was applied at 10 ng/ml, and RSV was applied at 10 nM in all the experiments. n = 6 per group ( a – f ) and n = 3 per group ( g – j ). The data represent the mean ± SD. * P

    Journal: Experimental & Molecular Medicine

    Article Title: Resveratrol enhances the functionality and improves the regeneration of mesenchymal stem cell aggregates

    doi: 10.1038/s12276-018-0109-y

    Figure Lengend Snippet: Resveratrol (RSV) rescued the cell aggregate formation and osteogenic differentiation of N-PDLSCs under inflammatory cytokine tumor necrosis factor alpha (TNF-α) treatment. a – c Histological analysis of PDLSC aggregates ( a ) showed that N-PDLSCs under TNF-α treatment formed thinner aggregates ( b ) with less collagen deposition ( c ), which could be rescued by RSV application. Bars: 50 μm. d Osteogenesis of PDLSC aggregates demonstrated by ALP (top) and alizarin red staining (bottom). Bars: 5 mm (top) and 50 μm (bottom). e , f RSV treatment restored the ALP activity ( e ) and mineralizing activity ( f ) of N-PDLSC aggregates under TNF-α application. g Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of the osteogenic marker genes Alp , Runt-related transcription factor 2 ( Runx2 ), Osteocalcin ( Ocn ), and Collagen I ( Col1 ) in PDLSC aggregates after osteogenic induction. The data showed that RSV rescued the osteogenic potential of N-PDLSC aggregates under TNF-α treatment. h Western blot analysis of the extracellular matrix (ECM) and osteogenic marker genes Col1, Periostin, and Runx2 in PDLSC aggregates. i Western blot analysis of molecular targets related to key signaling pathways in PDLSC aggregates: the Sirtuin 1 (Sirt1)-peroxisome proliferator-activated receptor gamma coactivator-1 alpha (Pgc1α) pathway; the nuclear factor kappaB (NFκB) pathway; and the adenosine monophosphate-activated protein kinase (AMPK) pathway. j Western blot analysis showed that RSV improved osteogenesis and ECM deposition in N-PDLSC aggregates under TNF-α treatment, possibly by inhibiting the NFκB pathway and stimulating the Pgc1α and AMPK pathways. TNF-α was applied at 10 ng/ml, and RSV was applied at 10 nM in all the experiments. n = 6 per group ( a – f ) and n = 3 per group ( g – j ). The data represent the mean ± SD. * P

    Article Snippet: The membranes were incubated overnight at 4 °C with the following antibodies: a rabbit anti-human Runx2 antibody (Santa Cruz Biotechnology) at a concentration of 1:1000, a mouse anti-human Col1 antibody (Abcam) at a concentration of 1:1000, a rabbit anti-human Periostin antibody (Abcam) at a concentration of 1:1000, a rabbit anti-human Sirtuin 1 (Sirt1) antibody (Abcam) at a concentration of 1:1000, a rabbit anti-human peroxisome proliferator-activated receptor gamma coactivator-1 alpha (Pgc1α) antibody (Abcam) at a concentration of 1:1000, a rabbit anti-human p -adenosine monophosphate-activated protein kinase (Ampk) antibody (Cell Signaling Technology) at a concentration of 1:1000, a mouse anti-human p-NFκB p65 antibody (Santa Cruz Biotechnology) at a concentration of 1:1000, a rabbit anti-human p-NFκB p65 antibody (Santa Cruz Biotechnology) at a concentration of 1:1000, and a mouse anti-human Gapdh antibody (Abcam) at a concentration of 1:4000.

    Techniques: ALP Assay, Staining, Activity Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Marker, Western Blot

    Effects of PGC-1α knockdown on the mitochondrial membrane potential in the HEC-1A cells. Notes: ( A ) Cells were stained with JC-1 and analyzed with flow cytometry after 72 hours of transfection with siRNA-2 or NC siRNA. siRNA-2 treatment increased the number of cells with low Δ ψ m . ( B ) Quantification of the results showed that siRNA-2 treatment increased the number of cells with low Δ ψ m . Results are shown as the mean ± SD values of three independent experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: Effects of PGC-1α knockdown on the mitochondrial membrane potential in the HEC-1A cells. Notes: ( A ) Cells were stained with JC-1 and analyzed with flow cytometry after 72 hours of transfection with siRNA-2 or NC siRNA. siRNA-2 treatment increased the number of cells with low Δ ψ m . ( B ) Quantification of the results showed that siRNA-2 treatment increased the number of cells with low Δ ψ m . Results are shown as the mean ± SD values of three independent experiments. * P

    Article Snippet: The primary antibody against human PGC-1α was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA), and the primary antibodies against human Bcl-2 and Bax were obtained from Zhongshan Biotechnology Co. (Beijing, People’s Republic of China).

    Techniques: Pyrolysis Gas Chromatography, Staining, Flow Cytometry, Cytometry, Transfection

    Knockdown of PGC-1α increased Bax expression and reduced Bcl-2 expression in the HEC-1A cells. Notes: ( A–C ) Western blot ratio analysis of Bax and β-actin, Bcl-2 and β-actin, and Bax and Bcl-2 is shown. In the siRNA-2 group, Bcl-2 expression was lower than in the NC-treated and untreated cells, whereas Bax expression was higher. Results are shown as the mean ± SD values of three independent experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: Knockdown of PGC-1α increased Bax expression and reduced Bcl-2 expression in the HEC-1A cells. Notes: ( A–C ) Western blot ratio analysis of Bax and β-actin, Bcl-2 and β-actin, and Bax and Bcl-2 is shown. In the siRNA-2 group, Bcl-2 expression was lower than in the NC-treated and untreated cells, whereas Bax expression was higher. Results are shown as the mean ± SD values of three independent experiments. * P

    Article Snippet: The primary antibody against human PGC-1α was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA), and the primary antibodies against human Bcl-2 and Bax were obtained from Zhongshan Biotechnology Co. (Beijing, People’s Republic of China).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Western Blot

    PGC-1α expression in the HEC-1A cells after 72 hours of transfection with siRNAs. Notes: ( A ) Real-time PCR analysis showed that all three siRNAs reduced the expression of PGC-1α after 72 hours of transfection compared with that in the NC siRNA-treated cells or in the untreated HEC-1A cells. siRNA-2 significantly reduced the expression of PGC-1α. ( B ) Western blot analysis showed that siRNA-2 reduced the expression of PGC-1α after 72 hours of transfection compared with that in the siRNA-transfected cells or in the untreated HEC-1A cells. Data are the mean ± SD values of three independent experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: PGC-1α expression in the HEC-1A cells after 72 hours of transfection with siRNAs. Notes: ( A ) Real-time PCR analysis showed that all three siRNAs reduced the expression of PGC-1α after 72 hours of transfection compared with that in the NC siRNA-treated cells or in the untreated HEC-1A cells. siRNA-2 significantly reduced the expression of PGC-1α. ( B ) Western blot analysis showed that siRNA-2 reduced the expression of PGC-1α after 72 hours of transfection compared with that in the siRNA-transfected cells or in the untreated HEC-1A cells. Data are the mean ± SD values of three independent experiments. * P

    Article Snippet: The primary antibody against human PGC-1α was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA), and the primary antibodies against human Bcl-2 and Bax were obtained from Zhongshan Biotechnology Co. (Beijing, People’s Republic of China).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Transfection, Real-time Polymerase Chain Reaction, Western Blot

    Effects of PGC-1α knockdown on the cell cycle in the HEC-1A cells. Notes: ( A ) cell-cycle distribution in the HEC-1A cells after 72 hours of transfection with siRNA-2 or NC, or in the untreated control cells. ( B ) Percentages of cells in the G1, S, and G2/M phases of the cell cycle (n=3). Values are given as the mean ± SD of triplicate experiments. P > 0.05 (ANOVA). Abbreviations: PGC-1α, peroxisome proliferator-activated receptor gamma coactivator-1 alpha; siRNA, small interfering RNA; NC, negative control.

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: Effects of PGC-1α knockdown on the cell cycle in the HEC-1A cells. Notes: ( A ) cell-cycle distribution in the HEC-1A cells after 72 hours of transfection with siRNA-2 or NC, or in the untreated control cells. ( B ) Percentages of cells in the G1, S, and G2/M phases of the cell cycle (n=3). Values are given as the mean ± SD of triplicate experiments. P > 0.05 (ANOVA). Abbreviations: PGC-1α, peroxisome proliferator-activated receptor gamma coactivator-1 alpha; siRNA, small interfering RNA; NC, negative control.

    Article Snippet: The primary antibody against human PGC-1α was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA), and the primary antibodies against human Bcl-2 and Bax were obtained from Zhongshan Biotechnology Co. (Beijing, People’s Republic of China).

    Techniques: Pyrolysis Gas Chromatography, Transfection, Small Interfering RNA, Negative Control

    Effects of PGC-1α knockdown on apoptosis of HEC-1A cells. Notes: ( A ) Cell apoptosis was detected with annexin V-FITC/PI double staining and flow cytometry. ( B ) Values (intensity of fluorescent positive cells during early apoptotic events) are given as mean ± SD of triplicate experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: Effects of PGC-1α knockdown on apoptosis of HEC-1A cells. Notes: ( A ) Cell apoptosis was detected with annexin V-FITC/PI double staining and flow cytometry. ( B ) Values (intensity of fluorescent positive cells during early apoptotic events) are given as mean ± SD of triplicate experiments. * P

    Article Snippet: The primary antibody against human PGC-1α was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA), and the primary antibodies against human Bcl-2 and Bax were obtained from Zhongshan Biotechnology Co. (Beijing, People’s Republic of China).

    Techniques: Pyrolysis Gas Chromatography, Double Staining, Flow Cytometry, Cytometry

    Immunofluorescence staining of apoptotic nuclei ( green ) and mitochondria ( red ) in human musculoskeletal cell lines after PGC-1α plasmid and TFAM siRNA co-transfection. (a) Nara-H, a human MFH cell line and (b) KTHOS, a human osteosarcoma cell line.

    Journal: Scientific Reports

    Article Title: Regulation of Mitochondrial Proliferation by PGC-1α Induces Cellular Apoptosis in Musculoskeletal Malignancies

    doi: 10.1038/srep03916

    Figure Lengend Snippet: Immunofluorescence staining of apoptotic nuclei ( green ) and mitochondria ( red ) in human musculoskeletal cell lines after PGC-1α plasmid and TFAM siRNA co-transfection. (a) Nara-H, a human MFH cell line and (b) KTHOS, a human osteosarcoma cell line.

    Article Snippet: Primary antibodies used in immunoblot analyses were: anti-human PGC-1α antibody (1:1000) (Cell Signaling Technology), anti-human TFAM antibody (1:1000) (Abnova), anti-human cleaved caspase-3 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-8 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-9 antibody (1:500) (Cell Signaling Technology), anti-human cytochrome c antibody (1:1000) (eBiosience Inc., San Diego, CA, USA), anti-human Bax antibody (1:1000) (Cell Signaling Technology) and anti-human α-tubulin antibody (1:10000) (Sigma-Aldrich).

    Techniques: Immunofluorescence, Staining, Pyrolysis Gas Chromatography, Plasmid Preparation, Cotransfection

    Effects of PGC-1α plasmid and TFAM siRNA co-transfection on mitochondrial biogenesis in a human MFH cell line, Nara-H. (a) Protein expression of PGC-1α and TFAM were evaluated by immunoblot analysis in Nara-H cells that were co-transfected with PGC-1α plasmid (or control plasmid) and TFAM siRNA (or control siRNA). (b) Relative mtDNA numbers in plasmid/siRNA transfected Nara-H cells were assessed by qRT-PCR. Data represent mean ± SE of at least three independent experiments (* P

    Journal: Scientific Reports

    Article Title: Regulation of Mitochondrial Proliferation by PGC-1α Induces Cellular Apoptosis in Musculoskeletal Malignancies

    doi: 10.1038/srep03916

    Figure Lengend Snippet: Effects of PGC-1α plasmid and TFAM siRNA co-transfection on mitochondrial biogenesis in a human MFH cell line, Nara-H. (a) Protein expression of PGC-1α and TFAM were evaluated by immunoblot analysis in Nara-H cells that were co-transfected with PGC-1α plasmid (or control plasmid) and TFAM siRNA (or control siRNA). (b) Relative mtDNA numbers in plasmid/siRNA transfected Nara-H cells were assessed by qRT-PCR. Data represent mean ± SE of at least three independent experiments (* P

    Article Snippet: Primary antibodies used in immunoblot analyses were: anti-human PGC-1α antibody (1:1000) (Cell Signaling Technology), anti-human TFAM antibody (1:1000) (Abnova), anti-human cleaved caspase-3 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-8 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-9 antibody (1:500) (Cell Signaling Technology), anti-human cytochrome c antibody (1:1000) (eBiosience Inc., San Diego, CA, USA), anti-human Bax antibody (1:1000) (Cell Signaling Technology) and anti-human α-tubulin antibody (1:10000) (Sigma-Aldrich).

    Techniques: Pyrolysis Gas Chromatography, Plasmid Preparation, Cotransfection, Expressing, Transfection, Quantitative RT-PCR

    Mitochondrial apoptotic activity after PGC-1α plasmid and TFAM siRNA co-transfection in human musculoskeletal cell lines. Apoptotic activity and mitochondrial numbers were assessed by flow cytometry in human MFH (Nara-H) (a) and human osteosarcoma (KTHOS) (b) cell lines. Results were normalized to control cell (control plasmid and control siRNA co-transfection) mean value. Data represent mean ± SE of at least three independent experiments (* P

    Journal: Scientific Reports

    Article Title: Regulation of Mitochondrial Proliferation by PGC-1α Induces Cellular Apoptosis in Musculoskeletal Malignancies

    doi: 10.1038/srep03916

    Figure Lengend Snippet: Mitochondrial apoptotic activity after PGC-1α plasmid and TFAM siRNA co-transfection in human musculoskeletal cell lines. Apoptotic activity and mitochondrial numbers were assessed by flow cytometry in human MFH (Nara-H) (a) and human osteosarcoma (KTHOS) (b) cell lines. Results were normalized to control cell (control plasmid and control siRNA co-transfection) mean value. Data represent mean ± SE of at least three independent experiments (* P

    Article Snippet: Primary antibodies used in immunoblot analyses were: anti-human PGC-1α antibody (1:1000) (Cell Signaling Technology), anti-human TFAM antibody (1:1000) (Abnova), anti-human cleaved caspase-3 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-8 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-9 antibody (1:500) (Cell Signaling Technology), anti-human cytochrome c antibody (1:1000) (eBiosience Inc., San Diego, CA, USA), anti-human Bax antibody (1:1000) (Cell Signaling Technology) and anti-human α-tubulin antibody (1:10000) (Sigma-Aldrich).

    Techniques: Activity Assay, Pyrolysis Gas Chromatography, Plasmid Preparation, Cotransfection, Flow Cytometry

    Imuunohistochemical staining for PGC-1α (a) and TFAM (b) in human musculoskeletal tumors and normal muscle tissue samples. Arrows indicate PGC-1α (a) and TFAM (b) stained cells.

    Journal: Scientific Reports

    Article Title: Regulation of Mitochondrial Proliferation by PGC-1α Induces Cellular Apoptosis in Musculoskeletal Malignancies

    doi: 10.1038/srep03916

    Figure Lengend Snippet: Imuunohistochemical staining for PGC-1α (a) and TFAM (b) in human musculoskeletal tumors and normal muscle tissue samples. Arrows indicate PGC-1α (a) and TFAM (b) stained cells.

    Article Snippet: Primary antibodies used in immunoblot analyses were: anti-human PGC-1α antibody (1:1000) (Cell Signaling Technology), anti-human TFAM antibody (1:1000) (Abnova), anti-human cleaved caspase-3 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-8 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-9 antibody (1:500) (Cell Signaling Technology), anti-human cytochrome c antibody (1:1000) (eBiosience Inc., San Diego, CA, USA), anti-human Bax antibody (1:1000) (Cell Signaling Technology) and anti-human α-tubulin antibody (1:10000) (Sigma-Aldrich).

    Techniques: Staining, Pyrolysis Gas Chromatography

    Western immunoblot analyses for apoptosis-related proteins in PGC-1α plasmid/TFAM siRNA transfected MFH cells. (a) Expression of cleaved forms of caspase-3, -8 and -9. (b) Expression of cytochrome c and Bax were separately evaluated in mitochondrial and cytoplasmic fractions.

    Journal: Scientific Reports

    Article Title: Regulation of Mitochondrial Proliferation by PGC-1α Induces Cellular Apoptosis in Musculoskeletal Malignancies

    doi: 10.1038/srep03916

    Figure Lengend Snippet: Western immunoblot analyses for apoptosis-related proteins in PGC-1α plasmid/TFAM siRNA transfected MFH cells. (a) Expression of cleaved forms of caspase-3, -8 and -9. (b) Expression of cytochrome c and Bax were separately evaluated in mitochondrial and cytoplasmic fractions.

    Article Snippet: Primary antibodies used in immunoblot analyses were: anti-human PGC-1α antibody (1:1000) (Cell Signaling Technology), anti-human TFAM antibody (1:1000) (Abnova), anti-human cleaved caspase-3 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-8 antibody (1:500) (Cell Signaling Technology), anti-human cleaved caspase-9 antibody (1:500) (Cell Signaling Technology), anti-human cytochrome c antibody (1:1000) (eBiosience Inc., San Diego, CA, USA), anti-human Bax antibody (1:1000) (Cell Signaling Technology) and anti-human α-tubulin antibody (1:10000) (Sigma-Aldrich).

    Techniques: Western Blot, Pyrolysis Gas Chromatography, Plasmid Preparation, Transfection, Expressing

    Palmitic acid induced type 2 diabetes-like features in HepG2 cells. (A) Lipid level was increased in PA concentration-dependent manner in PA-treated cells determined by Nile Red staining. (B) Lipid metabolism-related proteins, PPARα and PGC-1α, were reduced in PA-treated cells. (C) Transcription levels of PPARα and PGC-1α were decreased by PA treatment. (D) Western blot results show that preproinsulin levels increased, and IRS2 and GLUT2 expression decreased in PA-treated cells in a concentration-dependent manner. (E) Transcription levels of insulin elevated and of IRS2 and GLUT2 declined following PA treatment. The upper panel shows representative electrophorogram of the gene bands from RT-PCR and lower pane ls show the corresponding histograms from real-time qPCR in C and E. *p

    Journal: International Journal of Biological Sciences

    Article Title: Intracellular Insulin and Impaired Autophagy in a Zebrafish model and a Cell Model of Type 2 diabetes

    doi: 10.7150/ijbs.19249

    Figure Lengend Snippet: Palmitic acid induced type 2 diabetes-like features in HepG2 cells. (A) Lipid level was increased in PA concentration-dependent manner in PA-treated cells determined by Nile Red staining. (B) Lipid metabolism-related proteins, PPARα and PGC-1α, were reduced in PA-treated cells. (C) Transcription levels of PPARα and PGC-1α were decreased by PA treatment. (D) Western blot results show that preproinsulin levels increased, and IRS2 and GLUT2 expression decreased in PA-treated cells in a concentration-dependent manner. (E) Transcription levels of insulin elevated and of IRS2 and GLUT2 declined following PA treatment. The upper panel shows representative electrophorogram of the gene bands from RT-PCR and lower pane ls show the corresponding histograms from real-time qPCR in C and E. *p

    Article Snippet: The membranes were incubated overnight at 4°C with primary antibody including anti-human p62 polyclonal antibody (PM045, MBL, Nagoya, Japan) diluted 1:500, and anti-human β-ACTIN monoclonal antibody (TA-09, ZSGB-Bio, Beijing, China) diluted 1:1000, anti-human LC3B polyclonal antibody (PM036, MBL) diluted 1:500, anti-human LAMP2 monoclonal antibody (sc-5571, Santa Cruz Biotechnology, Dallas, TX) diluted 1:200, anti-human PPARA monoclonal antibody (AM8452b, Abgent, Suzhou, China) diluted 1:500, anti-human PGC-1α polyclonal antibody (ab54481, Abcam, Cambridge, UK) diluted 1:500, anti-human insulin polyclonal antibody (sc-7838,Santa Cruz Biotechnology) diluted 1:200, anti-human GLUT2 polyclonal antibody (sc-9117,Santa Cruz Biotechnology) diluted 1:200, and anti-human IRS2 polyclonal antibody (#4502, CST, Danvers, MA, USA) diluted 1:200.

    Techniques: Concentration Assay, Staining, Pyrolysis Gas Chromatography, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    Graphical summary of inter-variable connections. Figure 2 illustrates statistical connections of variables of interest. Bivariate correlations were calculated for the entire study population using the model of Spearman’s rho. R coefficients and p-values (p ≤ 0.05; in brackets) are presented in the grey box and summarised in a graphical model. For ease of reading, lifestyle factors have not been incorporated in detail, and are therefore only abstracted (bottom of figure). Abbreviations: UCB: unconjugated bilirubin; UGT1A1 : UGT1A1 genotype; pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1; Sirt-1: Sirtuin-1; FGF-21: Fibroblast growth factor 21; T3: Free triiodothyronine; BMI: Body mass index; LBM: Lean body mass; HbA1c: Glycated haemoglobin A1c; TChol: Total cholesterol; HDL: High density lipoprotein cholesterol; LDL: Low density lipoprotein cholesterol; TG: Triglyceride; LPA2: Lipoprotein A2; ApoA1: Apolipoprotein A1; ApoB: Apolipoprotein B.

    Journal: Scientific Reports

    Article Title: Features of an altered AMPK metabolic pathway in Gilbert’s Syndrome, and its role in metabolic health

    doi: 10.1038/srep30051

    Figure Lengend Snippet: Graphical summary of inter-variable connections. Figure 2 illustrates statistical connections of variables of interest. Bivariate correlations were calculated for the entire study population using the model of Spearman’s rho. R coefficients and p-values (p ≤ 0.05; in brackets) are presented in the grey box and summarised in a graphical model. For ease of reading, lifestyle factors have not been incorporated in detail, and are therefore only abstracted (bottom of figure). Abbreviations: UCB: unconjugated bilirubin; UGT1A1 : UGT1A1 genotype; pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1; Sirt-1: Sirtuin-1; FGF-21: Fibroblast growth factor 21; T3: Free triiodothyronine; BMI: Body mass index; LBM: Lean body mass; HbA1c: Glycated haemoglobin A1c; TChol: Total cholesterol; HDL: High density lipoprotein cholesterol; LDL: Low density lipoprotein cholesterol; TG: Triglyceride; LPA2: Lipoprotein A2; ApoA1: Apolipoprotein A1; ApoB: Apolipoprotein B.

    Article Snippet: The antibody set-up used was as follows: anti-phospho-AMPK: rabbit anti-human monoclonal to AMPK α1 (phos-T183) and AMPK α2 (phos-T172), (ab133448, Abcam); secondary antibody to phospho-AMPK: goat anti-rabbit IgG H & L AlexaFluor 488, (ab150077, Abcam); anti-PgC 1α: rabbit anti-human polyclonal to PgC 1α, PE-labelled, (orb124814, Biorbyt); anti-phospho-Ppar α: rabbit anti-human polyclonal to Ppar α (phos-Ser12), FITC-labelled, (bs-4055R-FITC, Bioss); anti-phospho-Ppar γ: rabbit anti-human polyclonal to Ppar γ (phos-Ser112), AlexaFluor 647 labelled, (bs-3737R-A647, Bioss).

    Techniques: Pyrolysis Gas Chromatography

    Comparison of measures of the AMPK pathway between the study groups (GS, C), including all subjects (male and female). Levels of (phosphorylated) proteins (AMPK α1/α2, Ppar α and γ, PgC 1α) were analysed using the method of flow cytometry. Data are expressed as relative fluorescence units [rfU], and compared between subjects with Gilbert’s syndrome (GS; UGT1A1 *28 promoter mutation), and controls (C). * Indicates significant differences between groups (p ≤ 0.05). Medians can be found in Table 2 . Abbreviations: pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1.

    Journal: Scientific Reports

    Article Title: Features of an altered AMPK metabolic pathway in Gilbert’s Syndrome, and its role in metabolic health

    doi: 10.1038/srep30051

    Figure Lengend Snippet: Comparison of measures of the AMPK pathway between the study groups (GS, C), including all subjects (male and female). Levels of (phosphorylated) proteins (AMPK α1/α2, Ppar α and γ, PgC 1α) were analysed using the method of flow cytometry. Data are expressed as relative fluorescence units [rfU], and compared between subjects with Gilbert’s syndrome (GS; UGT1A1 *28 promoter mutation), and controls (C). * Indicates significant differences between groups (p ≤ 0.05). Medians can be found in Table 2 . Abbreviations: pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1.

    Article Snippet: The antibody set-up used was as follows: anti-phospho-AMPK: rabbit anti-human monoclonal to AMPK α1 (phos-T183) and AMPK α2 (phos-T172), (ab133448, Abcam); secondary antibody to phospho-AMPK: goat anti-rabbit IgG H & L AlexaFluor 488, (ab150077, Abcam); anti-PgC 1α: rabbit anti-human polyclonal to PgC 1α, PE-labelled, (orb124814, Biorbyt); anti-phospho-Ppar α: rabbit anti-human polyclonal to Ppar α (phos-Ser12), FITC-labelled, (bs-4055R-FITC, Bioss); anti-phospho-Ppar γ: rabbit anti-human polyclonal to Ppar γ (phos-Ser112), AlexaFluor 647 labelled, (bs-3737R-A647, Bioss).

    Techniques: Pyrolysis Gas Chromatography, Flow Cytometry, Cytometry, Fluorescence, Mutagenesis

    Correlations of UCB ( a–d ) and the UGT1A1 genotype ( e–h ), with measures of the AMPK pathway. Figure 3 illustrates gender-specific correlations of UCB and the UGT1A1 genotype with measures of the AMPK pathway. Bivariate correlations between UCB/ UGT1A1 genotype (-TA repeats: 6/6 controls, 6/7 heterozygous, 7/7 Gilbert’s syndrome) and measures of the AMPK pathway were calculated for each gender (m = male, f = female), using the model of Spearman’s rho. R coefficients and p-values (p ≤ 0.05; in brackets) are as follows: UCB * pAMPK α1/α2: m 0.594 (0.000); f 0.255 (0.122), UCB * PgC1 α: m 0.376 (0.001); f 0.467 (0.003), UCB * pPpar α: m 0.435 (0.000); f 0.575 (0.000), UCB * pPpar γ: m 0.354 (0.001); f 0.324 (0.047), UGT1A1 * pAMPK α1/α2: m 0.541 (p 0.000); f 0.156 (p 0.362), UGT1A1 * PgC1 α: m 0.265 (p 0.023); f 0.551 (p 0.001), UGT1A1 * Ppar α: m 0.365 (p 0.002); f 0.661 (p 0.000), UGT1A1 * Ppar γ: m 0.191 (p 0.023); f 0.435 (p 0.008). Abbreviations: UCB: unconjugated bilirubin; pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1; WT: wild type (control subjects); GS: Gilbert’s syndrome.

    Journal: Scientific Reports

    Article Title: Features of an altered AMPK metabolic pathway in Gilbert’s Syndrome, and its role in metabolic health

    doi: 10.1038/srep30051

    Figure Lengend Snippet: Correlations of UCB ( a–d ) and the UGT1A1 genotype ( e–h ), with measures of the AMPK pathway. Figure 3 illustrates gender-specific correlations of UCB and the UGT1A1 genotype with measures of the AMPK pathway. Bivariate correlations between UCB/ UGT1A1 genotype (-TA repeats: 6/6 controls, 6/7 heterozygous, 7/7 Gilbert’s syndrome) and measures of the AMPK pathway were calculated for each gender (m = male, f = female), using the model of Spearman’s rho. R coefficients and p-values (p ≤ 0.05; in brackets) are as follows: UCB * pAMPK α1/α2: m 0.594 (0.000); f 0.255 (0.122), UCB * PgC1 α: m 0.376 (0.001); f 0.467 (0.003), UCB * pPpar α: m 0.435 (0.000); f 0.575 (0.000), UCB * pPpar γ: m 0.354 (0.001); f 0.324 (0.047), UGT1A1 * pAMPK α1/α2: m 0.541 (p 0.000); f 0.156 (p 0.362), UGT1A1 * PgC1 α: m 0.265 (p 0.023); f 0.551 (p 0.001), UGT1A1 * Ppar α: m 0.365 (p 0.002); f 0.661 (p 0.000), UGT1A1 * Ppar γ: m 0.191 (p 0.023); f 0.435 (p 0.008). Abbreviations: UCB: unconjugated bilirubin; pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1; WT: wild type (control subjects); GS: Gilbert’s syndrome.

    Article Snippet: The antibody set-up used was as follows: anti-phospho-AMPK: rabbit anti-human monoclonal to AMPK α1 (phos-T183) and AMPK α2 (phos-T172), (ab133448, Abcam); secondary antibody to phospho-AMPK: goat anti-rabbit IgG H & L AlexaFluor 488, (ab150077, Abcam); anti-PgC 1α: rabbit anti-human polyclonal to PgC 1α, PE-labelled, (orb124814, Biorbyt); anti-phospho-Ppar α: rabbit anti-human polyclonal to Ppar α (phos-Ser12), FITC-labelled, (bs-4055R-FITC, Bioss); anti-phospho-Ppar γ: rabbit anti-human polyclonal to Ppar γ (phos-Ser112), AlexaFluor 647 labelled, (bs-3737R-A647, Bioss).

    Techniques: Pyrolysis Gas Chromatography

    Abstract of inter-variable connection and dependence, based on regression analysis. Stepwise linear regression analysis was performed to assess inter-variable dependence and explanatory power (%), based on corrected R 2 values and a p-value of ≤0.05 ( Tables 5 and 6 ). C effect found in control subjects only, GS effect found in GS subjects only, (All remaining unspecified correlations are valid for both study groups.), Abbreviations: UCB: unconjugated bilirubin; UGT1A1 : UGT1A1 genotype; AMPKα1: 5′-AMP activated kinase α1 gene expression; pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1; Sirt-1: Sirtuin-1; BMI: Body mass index; LBM: Lean body mass.

    Journal: Scientific Reports

    Article Title: Features of an altered AMPK metabolic pathway in Gilbert’s Syndrome, and its role in metabolic health

    doi: 10.1038/srep30051

    Figure Lengend Snippet: Abstract of inter-variable connection and dependence, based on regression analysis. Stepwise linear regression analysis was performed to assess inter-variable dependence and explanatory power (%), based on corrected R 2 values and a p-value of ≤0.05 ( Tables 5 and 6 ). C effect found in control subjects only, GS effect found in GS subjects only, (All remaining unspecified correlations are valid for both study groups.), Abbreviations: UCB: unconjugated bilirubin; UGT1A1 : UGT1A1 genotype; AMPKα1: 5′-AMP activated kinase α1 gene expression; pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1; Sirt-1: Sirtuin-1; BMI: Body mass index; LBM: Lean body mass.

    Article Snippet: The antibody set-up used was as follows: anti-phospho-AMPK: rabbit anti-human monoclonal to AMPK α1 (phos-T183) and AMPK α2 (phos-T172), (ab133448, Abcam); secondary antibody to phospho-AMPK: goat anti-rabbit IgG H & L AlexaFluor 488, (ab150077, Abcam); anti-PgC 1α: rabbit anti-human polyclonal to PgC 1α, PE-labelled, (orb124814, Biorbyt); anti-phospho-Ppar α: rabbit anti-human polyclonal to Ppar α (phos-Ser12), FITC-labelled, (bs-4055R-FITC, Bioss); anti-phospho-Ppar γ: rabbit anti-human polyclonal to Ppar γ (phos-Ser112), AlexaFluor 647 labelled, (bs-3737R-A647, Bioss).

    Techniques: Expressing, Pyrolysis Gas Chromatography

    Overexpression of pgc-1α suppresses 4-ClBQ induced down-regulation of catalase expression in HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure PGC-1α (A) mRNA and (B) protein levels in vector-alone and PGC-1α

    Journal: Toxicology in vitro : an international journal published in association with BIBRA

    Article Title: Down-regulation of peroxisome proliferator activated receptor γ coactivator 1α induces oxidative stress and toxicity of 1-(4-Chlorophenyl)-benzo-2,5-quinone in HaCaT human keratinocytes

    doi: 10.1016/j.tiv.2015.05.009

    Figure Lengend Snippet: Overexpression of pgc-1α suppresses 4-ClBQ induced down-regulation of catalase expression in HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure PGC-1α (A) mRNA and (B) protein levels in vector-alone and PGC-1α

    Article Snippet: Human pgc-1α was transiently overexpressed in HaCaT cells using human PGC-1α plasmid DNA (Addgene, plasmid No. 10974; Cambridge, MA).

    Techniques: Over Expression, Pyrolysis Gas Chromatography, Expressing, Quantitative RT-PCR, Plasmid Preparation

    pgc-1α expression is inhibited in 4-ClBQ treated HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure pgc-1α (A) mRNA and (B) protein levels in control and 4-ClBQ treated cells at 24 h after the addition of 4-ClBQ.

    Journal: Toxicology in vitro : an international journal published in association with BIBRA

    Article Title: Down-regulation of peroxisome proliferator activated receptor γ coactivator 1α induces oxidative stress and toxicity of 1-(4-Chlorophenyl)-benzo-2,5-quinone in HaCaT human keratinocytes

    doi: 10.1016/j.tiv.2015.05.009

    Figure Lengend Snippet: pgc-1α expression is inhibited in 4-ClBQ treated HaCaT cells. Quantitative RT-PCR and immunoblotting assays were used to measure pgc-1α (A) mRNA and (B) protein levels in control and 4-ClBQ treated cells at 24 h after the addition of 4-ClBQ.

    Article Snippet: Human pgc-1α was transiently overexpressed in HaCaT cells using human PGC-1α plasmid DNA (Addgene, plasmid No. 10974; Cambridge, MA).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Quantitative RT-PCR

    Overexpression of pgc-1α suppresses 4-ClBQ induced oxidative stress and toxicity in HaCaT cells. Flow cytometry measurements of MitoSOX Red oxidation were used to determine cellular ROS levels in control and 4-ClBQ treated vector-alone and pgc-1α

    Journal: Toxicology in vitro : an international journal published in association with BIBRA

    Article Title: Down-regulation of peroxisome proliferator activated receptor γ coactivator 1α induces oxidative stress and toxicity of 1-(4-Chlorophenyl)-benzo-2,5-quinone in HaCaT human keratinocytes

    doi: 10.1016/j.tiv.2015.05.009

    Figure Lengend Snippet: Overexpression of pgc-1α suppresses 4-ClBQ induced oxidative stress and toxicity in HaCaT cells. Flow cytometry measurements of MitoSOX Red oxidation were used to determine cellular ROS levels in control and 4-ClBQ treated vector-alone and pgc-1α

    Article Snippet: Human pgc-1α was transiently overexpressed in HaCaT cells using human PGC-1α plasmid DNA (Addgene, plasmid No. 10974; Cambridge, MA).

    Techniques: Over Expression, Pyrolysis Gas Chromatography, Flow Cytometry, Cytometry, Plasmid Preparation

    Effects of PGC-1α knockdown on the mitochondrial membrane potential in the HEC-1A cells. Notes: ( A ) Cells were stained with JC-1 and analyzed with flow cytometry after 72 hours of transfection with siRNA-2 or NC siRNA. siRNA-2 treatment increased the number of cells with low Δ ψ m . ( B ) Quantification of the results showed that siRNA-2 treatment increased the number of cells with low Δ ψ m . Results are shown as the mean ± SD values of three independent experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: Effects of PGC-1α knockdown on the mitochondrial membrane potential in the HEC-1A cells. Notes: ( A ) Cells were stained with JC-1 and analyzed with flow cytometry after 72 hours of transfection with siRNA-2 or NC siRNA. siRNA-2 treatment increased the number of cells with low Δ ψ m . ( B ) Quantification of the results showed that siRNA-2 treatment increased the number of cells with low Δ ψ m . Results are shown as the mean ± SD values of three independent experiments. * P

    Article Snippet: Transfection Three small interfering RNAs (siRNAs) targeting different sites in the human PGC-1α mRNA (GenBank accession no NM_013261.3) were designed and synthesized by GenePharma Co., Ltd (Jiangsu, People’s Republic of China) to knock down PGC-1α expression, and a control siRNA that did not target PGC-1α mRNA was synthesized as the negative control (NC).

    Techniques: Pyrolysis Gas Chromatography, Staining, Flow Cytometry, Cytometry, Transfection

    Knockdown of PGC-1α increased Bax expression and reduced Bcl-2 expression in the HEC-1A cells. Notes: ( A–C ) Western blot ratio analysis of Bax and β-actin, Bcl-2 and β-actin, and Bax and Bcl-2 is shown. In the siRNA-2 group, Bcl-2 expression was lower than in the NC-treated and untreated cells, whereas Bax expression was higher. Results are shown as the mean ± SD values of three independent experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: Knockdown of PGC-1α increased Bax expression and reduced Bcl-2 expression in the HEC-1A cells. Notes: ( A–C ) Western blot ratio analysis of Bax and β-actin, Bcl-2 and β-actin, and Bax and Bcl-2 is shown. In the siRNA-2 group, Bcl-2 expression was lower than in the NC-treated and untreated cells, whereas Bax expression was higher. Results are shown as the mean ± SD values of three independent experiments. * P

    Article Snippet: Transfection Three small interfering RNAs (siRNAs) targeting different sites in the human PGC-1α mRNA (GenBank accession no NM_013261.3) were designed and synthesized by GenePharma Co., Ltd (Jiangsu, People’s Republic of China) to knock down PGC-1α expression, and a control siRNA that did not target PGC-1α mRNA was synthesized as the negative control (NC).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Western Blot

    PGC-1α expression in the HEC-1A cells after 72 hours of transfection with siRNAs. Notes: ( A ) Real-time PCR analysis showed that all three siRNAs reduced the expression of PGC-1α after 72 hours of transfection compared with that in the NC siRNA-treated cells or in the untreated HEC-1A cells. siRNA-2 significantly reduced the expression of PGC-1α. ( B ) Western blot analysis showed that siRNA-2 reduced the expression of PGC-1α after 72 hours of transfection compared with that in the siRNA-transfected cells or in the untreated HEC-1A cells. Data are the mean ± SD values of three independent experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: PGC-1α expression in the HEC-1A cells after 72 hours of transfection with siRNAs. Notes: ( A ) Real-time PCR analysis showed that all three siRNAs reduced the expression of PGC-1α after 72 hours of transfection compared with that in the NC siRNA-treated cells or in the untreated HEC-1A cells. siRNA-2 significantly reduced the expression of PGC-1α. ( B ) Western blot analysis showed that siRNA-2 reduced the expression of PGC-1α after 72 hours of transfection compared with that in the siRNA-transfected cells or in the untreated HEC-1A cells. Data are the mean ± SD values of three independent experiments. * P

    Article Snippet: Transfection Three small interfering RNAs (siRNAs) targeting different sites in the human PGC-1α mRNA (GenBank accession no NM_013261.3) were designed and synthesized by GenePharma Co., Ltd (Jiangsu, People’s Republic of China) to knock down PGC-1α expression, and a control siRNA that did not target PGC-1α mRNA was synthesized as the negative control (NC).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Transfection, Real-time Polymerase Chain Reaction, Western Blot

    Effects of PGC-1α knockdown on the cell cycle in the HEC-1A cells. Notes: ( A ) cell-cycle distribution in the HEC-1A cells after 72 hours of transfection with siRNA-2 or NC, or in the untreated control cells. ( B ) Percentages of cells in the G1, S, and G2/M phases of the cell cycle (n=3). Values are given as the mean ± SD of triplicate experiments. P > 0.05 (ANOVA). Abbreviations: PGC-1α, peroxisome proliferator-activated receptor gamma coactivator-1 alpha; siRNA, small interfering RNA; NC, negative control.

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: Effects of PGC-1α knockdown on the cell cycle in the HEC-1A cells. Notes: ( A ) cell-cycle distribution in the HEC-1A cells after 72 hours of transfection with siRNA-2 or NC, or in the untreated control cells. ( B ) Percentages of cells in the G1, S, and G2/M phases of the cell cycle (n=3). Values are given as the mean ± SD of triplicate experiments. P > 0.05 (ANOVA). Abbreviations: PGC-1α, peroxisome proliferator-activated receptor gamma coactivator-1 alpha; siRNA, small interfering RNA; NC, negative control.

    Article Snippet: Transfection Three small interfering RNAs (siRNAs) targeting different sites in the human PGC-1α mRNA (GenBank accession no NM_013261.3) were designed and synthesized by GenePharma Co., Ltd (Jiangsu, People’s Republic of China) to knock down PGC-1α expression, and a control siRNA that did not target PGC-1α mRNA was synthesized as the negative control (NC).

    Techniques: Pyrolysis Gas Chromatography, Transfection, Small Interfering RNA, Negative Control

    Effects of PGC-1α knockdown on apoptosis of HEC-1A cells. Notes: ( A ) Cell apoptosis was detected with annexin V-FITC/PI double staining and flow cytometry. ( B ) Values (intensity of fluorescent positive cells during early apoptotic events) are given as mean ± SD of triplicate experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Knockdown of peroxisome proliferator-activated receptor gamma coactivator-1 alpha increased apoptosis of human endometrial cancer HEC-1A cells

    doi: 10.2147/OTT.S102816

    Figure Lengend Snippet: Effects of PGC-1α knockdown on apoptosis of HEC-1A cells. Notes: ( A ) Cell apoptosis was detected with annexin V-FITC/PI double staining and flow cytometry. ( B ) Values (intensity of fluorescent positive cells during early apoptotic events) are given as mean ± SD of triplicate experiments. * P

    Article Snippet: Transfection Three small interfering RNAs (siRNAs) targeting different sites in the human PGC-1α mRNA (GenBank accession no NM_013261.3) were designed and synthesized by GenePharma Co., Ltd (Jiangsu, People’s Republic of China) to knock down PGC-1α expression, and a control siRNA that did not target PGC-1α mRNA was synthesized as the negative control (NC).

    Techniques: Pyrolysis Gas Chromatography, Double Staining, Flow Cytometry, Cytometry

    Anti-apoptotic effect of PGC-1α. Stable cells were treated with 0.5 mM H 2 O 2 for 6 h. ( A ) The expression bands of apoptotic proteins in Mock and PGC-1α-stable cells were compared via western blotting. Each bar graph represents the expression of PGC-1α ( B ), ratio of phosphorylated p53 at Ser 15 to total p53 ( C ), the level of activated caspase 3 (ratio of cleaved caspase 3 to caspase 3 ( D ), and the level of cytochrome C release from mitochondria to cytosol ( E ). β-actin levels were analyzed as internal controls. GAPDH and complex V α were used as internal controls in cytosol and in mitochondria fraction, respectively. Full-length blots of each tested protein are reported in Supplementary Figure S4 . Error bars denote the mean ± S.D. of triplicate samples. * p

    Journal: Scientific Reports

    Article Title: PGC-1α attenuates hydrogen peroxide-induced apoptotic cell death by upregulating Nrf-2 via GSK3β inactivation mediated by activated p38 in HK-2 Cells

    doi: 10.1038/s41598-017-04593-w

    Figure Lengend Snippet: Anti-apoptotic effect of PGC-1α. Stable cells were treated with 0.5 mM H 2 O 2 for 6 h. ( A ) The expression bands of apoptotic proteins in Mock and PGC-1α-stable cells were compared via western blotting. Each bar graph represents the expression of PGC-1α ( B ), ratio of phosphorylated p53 at Ser 15 to total p53 ( C ), the level of activated caspase 3 (ratio of cleaved caspase 3 to caspase 3 ( D ), and the level of cytochrome C release from mitochondria to cytosol ( E ). β-actin levels were analyzed as internal controls. GAPDH and complex V α were used as internal controls in cytosol and in mitochondria fraction, respectively. Full-length blots of each tested protein are reported in Supplementary Figure S4 . Error bars denote the mean ± S.D. of triplicate samples. * p

    Article Snippet: For over-expression of human PGC-1α in HK-2 cells, human PGC-1α/pCDNA4 plasmid was purchased from Addgene (Cat#10974, Cambridge, USA).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Western Blot

    Downregulation of PGC-1α in H 2 O 2 -treated HK-2 cells. To mimic I/R-induced oxidative stress in an in vitro system, we treated with H 2 O 2 in HK-2 cells. ( A ) Dose-dependent PGC-1α expression. HK-2 cells were treated with an indicated H 2 O 2 concentration (0, 0.2, 0.5, 1, and 2) for 6 h. ( B ) Time-dependent PGC-1α expression. HK-2 cells were treated with 0.5 mM H 2 O 2 for an indicated time (0, 3, 6, 12, and 24 h). ( C ) To assess the effect ROS in H 2 O 2 induced PGC-1α downregulation, cells were incubated for 6 h with 0.5 mM H 2 O 2 in the presence or absence of 20 mM NAC. The bar graph shows the relative protein expression of PGC-1α measured by densitometry. β-actin levels were analyzed as internal controls. Full-length blots of each tested protein are reported in Supplementary Figure S2 . Error bars denote the mean ± S.D. of triplicate samples. * p

    Journal: Scientific Reports

    Article Title: PGC-1α attenuates hydrogen peroxide-induced apoptotic cell death by upregulating Nrf-2 via GSK3β inactivation mediated by activated p38 in HK-2 Cells

    doi: 10.1038/s41598-017-04593-w

    Figure Lengend Snippet: Downregulation of PGC-1α in H 2 O 2 -treated HK-2 cells. To mimic I/R-induced oxidative stress in an in vitro system, we treated with H 2 O 2 in HK-2 cells. ( A ) Dose-dependent PGC-1α expression. HK-2 cells were treated with an indicated H 2 O 2 concentration (0, 0.2, 0.5, 1, and 2) for 6 h. ( B ) Time-dependent PGC-1α expression. HK-2 cells were treated with 0.5 mM H 2 O 2 for an indicated time (0, 3, 6, 12, and 24 h). ( C ) To assess the effect ROS in H 2 O 2 induced PGC-1α downregulation, cells were incubated for 6 h with 0.5 mM H 2 O 2 in the presence or absence of 20 mM NAC. The bar graph shows the relative protein expression of PGC-1α measured by densitometry. β-actin levels were analyzed as internal controls. Full-length blots of each tested protein are reported in Supplementary Figure S2 . Error bars denote the mean ± S.D. of triplicate samples. * p

    Article Snippet: For over-expression of human PGC-1α in HK-2 cells, human PGC-1α/pCDNA4 plasmid was purchased from Addgene (Cat#10974, Cambridge, USA).

    Techniques: Pyrolysis Gas Chromatography, In Vitro, Expressing, Concentration Assay, Incubation

    Nrf-2 specific-protective effects by PGC-1α. To prove the dependence of Nrf-2 in anti-apoptotic and anti-oxidative effect of PGC-1α, the level of activated caspase 3. Full-length blots of each tested protein are reported in Supplementary Figure S6 ( A ) and the level of DCF fluorescence ( B ) were assessed in Nrf-2 suppressed PGC-1α cells under H 2 O 2 treatment, as earlier mentioned. Magnification at x200, Bar = 100 μm. Relative protein level and ROS level were expressed as fold normalized to the untreated Mock cells. β-actin levels were analyzed as internal controls. Error bars denote the mean ± S.D. of triplicate samples. * p

    Journal: Scientific Reports

    Article Title: PGC-1α attenuates hydrogen peroxide-induced apoptotic cell death by upregulating Nrf-2 via GSK3β inactivation mediated by activated p38 in HK-2 Cells

    doi: 10.1038/s41598-017-04593-w

    Figure Lengend Snippet: Nrf-2 specific-protective effects by PGC-1α. To prove the dependence of Nrf-2 in anti-apoptotic and anti-oxidative effect of PGC-1α, the level of activated caspase 3. Full-length blots of each tested protein are reported in Supplementary Figure S6 ( A ) and the level of DCF fluorescence ( B ) were assessed in Nrf-2 suppressed PGC-1α cells under H 2 O 2 treatment, as earlier mentioned. Magnification at x200, Bar = 100 μm. Relative protein level and ROS level were expressed as fold normalized to the untreated Mock cells. β-actin levels were analyzed as internal controls. Error bars denote the mean ± S.D. of triplicate samples. * p

    Article Snippet: For over-expression of human PGC-1α in HK-2 cells, human PGC-1α/pCDNA4 plasmid was purchased from Addgene (Cat#10974, Cambridge, USA).

    Techniques: Pyrolysis Gas Chromatography, Fluorescence

    The involvement of p38/GSK3β/Nrf-2 axis for cytoprotective effects of PGC-1α. To understand the molecular mechanism for cytoprotective effects of PGC-1α, the activation of Keap1-, GSK3β- or Hrd1 ( A ) and MAPKs ( B ), as an upstream signal molecules of Nrf-2, were compared to Mock and PGC-1α cells for indicated time points after H 2 O 2 treatment. To further elucidate the specificity of p38-mediated signal cascade for cytoprotective effects of PGC-1α cells, p38 specific effects were analyzed by western blotting ( C ) and MTT assay ( D ) in PGC-1α cells treated with H 2 O 2 in the presence or absence of p38 inhibitor (SB203580) for 2 h or 4 h, respectively. Inhibitor of ERK1/2 (PD98059) was used as non-effective control on PGC-1α effect. Activation of p38 or inactivation of GSK3β was checked with phosphor specific antibody at Thr180/Tyr182 residue or at Ser9 residue, respectively. Full-length blots of each tested protein are reported in Supplementary Figure S7 . Error bars denote the mean ± S.D. of triplicate samples. MTT assay was performed to n = 7. * p

    Journal: Scientific Reports

    Article Title: PGC-1α attenuates hydrogen peroxide-induced apoptotic cell death by upregulating Nrf-2 via GSK3β inactivation mediated by activated p38 in HK-2 Cells

    doi: 10.1038/s41598-017-04593-w

    Figure Lengend Snippet: The involvement of p38/GSK3β/Nrf-2 axis for cytoprotective effects of PGC-1α. To understand the molecular mechanism for cytoprotective effects of PGC-1α, the activation of Keap1-, GSK3β- or Hrd1 ( A ) and MAPKs ( B ), as an upstream signal molecules of Nrf-2, were compared to Mock and PGC-1α cells for indicated time points after H 2 O 2 treatment. To further elucidate the specificity of p38-mediated signal cascade for cytoprotective effects of PGC-1α cells, p38 specific effects were analyzed by western blotting ( C ) and MTT assay ( D ) in PGC-1α cells treated with H 2 O 2 in the presence or absence of p38 inhibitor (SB203580) for 2 h or 4 h, respectively. Inhibitor of ERK1/2 (PD98059) was used as non-effective control on PGC-1α effect. Activation of p38 or inactivation of GSK3β was checked with phosphor specific antibody at Thr180/Tyr182 residue or at Ser9 residue, respectively. Full-length blots of each tested protein are reported in Supplementary Figure S7 . Error bars denote the mean ± S.D. of triplicate samples. MTT assay was performed to n = 7. * p

    Article Snippet: For over-expression of human PGC-1α in HK-2 cells, human PGC-1α/pCDNA4 plasmid was purchased from Addgene (Cat#10974, Cambridge, USA).

    Techniques: Pyrolysis Gas Chromatography, Activation Assay, Western Blot, MTT Assay

    Protective effect of PGC-1α in H 2 O 2 -mediated cellular injury. To verify the role of PGC-1α in H 2 O 2 -mediated cellular injury, we developed stable cell lines over-expressing PGC-1α in HK-2 cells, as mentioned in the Materials and Methods section. ( A ) Expression of PGC-1α in Mock and PGC-1α stable cells. The protein expression (upper panel) of c-terminal c-Myc tagged PGC-1α was assessed with anti-c-Myc and stable cells were selected via the confirmation of zeocine expression (upper panel), which was contained in the backbone plasmid, pCDNA4 . Full-length blots of each tested protein are reported in Supplementary Figure S3 . ( B ) Cell viability of Mock and PGC-1α-stable cells. Stable cells were treated with 0.5 mM H 2 O 2 for an indicated time (0, 2, 4, and 6 h). Cell viability was evaluated by the MTT method. ( C ) Quantification of apoptotic cells. Mock and PGC-1α cells were treated with 0.5 mM H 2 O 2 for 4 h. Apoptotic cell numbers were measured by counting Annexin V positive cells using a FACSCalibur TM flow cytometry. ( D ) Apoptotic body formation in H 2 O 2 -treated Mock and PGC-1α stable cells. Apoptotic body formation (arrows), as an indicator of apoptosis, was determined by DAPI staining then photographing cells under fluorescence microscopy as described in the Materials and Methods. Image was magnified at x800, Bar = 20 μm. Error bars denote the mean ± S.D. of triplicate samples. * p

    Journal: Scientific Reports

    Article Title: PGC-1α attenuates hydrogen peroxide-induced apoptotic cell death by upregulating Nrf-2 via GSK3β inactivation mediated by activated p38 in HK-2 Cells

    doi: 10.1038/s41598-017-04593-w

    Figure Lengend Snippet: Protective effect of PGC-1α in H 2 O 2 -mediated cellular injury. To verify the role of PGC-1α in H 2 O 2 -mediated cellular injury, we developed stable cell lines over-expressing PGC-1α in HK-2 cells, as mentioned in the Materials and Methods section. ( A ) Expression of PGC-1α in Mock and PGC-1α stable cells. The protein expression (upper panel) of c-terminal c-Myc tagged PGC-1α was assessed with anti-c-Myc and stable cells were selected via the confirmation of zeocine expression (upper panel), which was contained in the backbone plasmid, pCDNA4 . Full-length blots of each tested protein are reported in Supplementary Figure S3 . ( B ) Cell viability of Mock and PGC-1α-stable cells. Stable cells were treated with 0.5 mM H 2 O 2 for an indicated time (0, 2, 4, and 6 h). Cell viability was evaluated by the MTT method. ( C ) Quantification of apoptotic cells. Mock and PGC-1α cells were treated with 0.5 mM H 2 O 2 for 4 h. Apoptotic cell numbers were measured by counting Annexin V positive cells using a FACSCalibur TM flow cytometry. ( D ) Apoptotic body formation in H 2 O 2 -treated Mock and PGC-1α stable cells. Apoptotic body formation (arrows), as an indicator of apoptosis, was determined by DAPI staining then photographing cells under fluorescence microscopy as described in the Materials and Methods. Image was magnified at x800, Bar = 20 μm. Error bars denote the mean ± S.D. of triplicate samples. * p

    Article Snippet: For over-expression of human PGC-1α in HK-2 cells, human PGC-1α/pCDNA4 plasmid was purchased from Addgene (Cat#10974, Cambridge, USA).

    Techniques: Pyrolysis Gas Chromatography, Stable Transfection, Expressing, Plasmid Preparation, MTT Assay, Flow Cytometry, Cytometry, Staining, Fluorescence, Microscopy

    Reduction of intracellular ROS by PGC-1α. To check the efficiency of PGC-1α overexpression in reducing ROS, stable cells were plated onto four well-cell culture slides (5 × 10 4 /well) and treated with 0.5 mM H 2 O 2 for 30 min. Cytosolic (magnification at x200, Bar = 100 μm) ( A ) and mitochondrial ROS (magnification at x400, Bar = 50 μm) ( B ) were labeled using CM-H 2 DCFDA or CM-H 2 XROS probes, respectively. Images were immediately visualized using confocal microscopy on a laser scanning microscope (LSM 510; Carl Zeiss), and analyzed using a LMS 5 browser imaging software. Error bars denote the mean ± S.D. of triplicate samples. * p

    Journal: Scientific Reports

    Article Title: PGC-1α attenuates hydrogen peroxide-induced apoptotic cell death by upregulating Nrf-2 via GSK3β inactivation mediated by activated p38 in HK-2 Cells

    doi: 10.1038/s41598-017-04593-w

    Figure Lengend Snippet: Reduction of intracellular ROS by PGC-1α. To check the efficiency of PGC-1α overexpression in reducing ROS, stable cells were plated onto four well-cell culture slides (5 × 10 4 /well) and treated with 0.5 mM H 2 O 2 for 30 min. Cytosolic (magnification at x200, Bar = 100 μm) ( A ) and mitochondrial ROS (magnification at x400, Bar = 50 μm) ( B ) were labeled using CM-H 2 DCFDA or CM-H 2 XROS probes, respectively. Images were immediately visualized using confocal microscopy on a laser scanning microscope (LSM 510; Carl Zeiss), and analyzed using a LMS 5 browser imaging software. Error bars denote the mean ± S.D. of triplicate samples. * p

    Article Snippet: For over-expression of human PGC-1α in HK-2 cells, human PGC-1α/pCDNA4 plasmid was purchased from Addgene (Cat#10974, Cambridge, USA).

    Techniques: Pyrolysis Gas Chromatography, Over Expression, Cell Culture, Labeling, Confocal Microscopy, Laser-Scanning Microscopy, Imaging, Software

    Downregulation of PGC-1α in I/R-induced kidney injury. ( A ) Plasma creatinin and blood urea nitrogen (BUN) levels of control vs. I/R-induced kidney injury in mice. ( B ) Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining of the kidney. Magnification at x100, Bar = 100 μm. ( C ) I/R-induced kidney injury was assessed by evaluating the expression of apoptotic proteins (ratio of Bax to Bcl2 and cleaved caspase 3 to caspase 3). ( D ) The mRNA expression of PGC-1α in control mice vs. those that underwent I/R-induced kidney injury. ( E ) Protein expression of PGC-1α in control mice vs. those that underwent I/R-induced kidney injury. The bar graph shows the indicated target expression measured by densitometry. GAPDH and β-actin levels were analyzed as internal controls. Full-length blots of each tested protein are reported in Supplementary Figure S1 . * p

    Journal: Scientific Reports

    Article Title: PGC-1α attenuates hydrogen peroxide-induced apoptotic cell death by upregulating Nrf-2 via GSK3β inactivation mediated by activated p38 in HK-2 Cells

    doi: 10.1038/s41598-017-04593-w

    Figure Lengend Snippet: Downregulation of PGC-1α in I/R-induced kidney injury. ( A ) Plasma creatinin and blood urea nitrogen (BUN) levels of control vs. I/R-induced kidney injury in mice. ( B ) Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining of the kidney. Magnification at x100, Bar = 100 μm. ( C ) I/R-induced kidney injury was assessed by evaluating the expression of apoptotic proteins (ratio of Bax to Bcl2 and cleaved caspase 3 to caspase 3). ( D ) The mRNA expression of PGC-1α in control mice vs. those that underwent I/R-induced kidney injury. ( E ) Protein expression of PGC-1α in control mice vs. those that underwent I/R-induced kidney injury. The bar graph shows the indicated target expression measured by densitometry. GAPDH and β-actin levels were analyzed as internal controls. Full-length blots of each tested protein are reported in Supplementary Figure S1 . * p

    Article Snippet: For over-expression of human PGC-1α in HK-2 cells, human PGC-1α/pCDNA4 plasmid was purchased from Addgene (Cat#10974, Cambridge, USA).

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, End Labeling, TUNEL Assay, Staining, Expressing

    Upregulation of Nrf-2 by PGC-1α overexpression. The change in Nrf-2 mRNA and protein levels by PGC-1α overexpression was evaluated in stable cells. Cells were treated with 0.5 mM H 2 O 2 for 6 h. The mRNA levels of PGC-1α ( A ) and Nrf-2 ( B ) were analyzed by real-time PCR with each PCR primer pair. GAPDH was analyzed as an internal control. ( C ) Upregulation of Nrf-2 protein in cytosol and nucleus fraction by PGC-1α overexpression. ( D ) Regulation of Nrf-2 mediated HO-1 expression by PGC-1α. To assess the involvement between PGC-1α expression and Nrf-2 mediated HO-1 (as a one of well-known downstream molecules activated by Nrf-2) expression for protective effect of PGC-1α, PGC-1α stable cells were incubated with Nrf-2 specific siRNA (30 and 50 nM), and then, after 2 days, cells were treated with 0.5 mM H 2 O 2 for 6 h. The protein level of Nrf-2 and HO-1 were determined by immunoblotting with specific antibodies. Relative protein level was expressed as fold increases normalized to the untreated Mock cells. β-actin levels were analyzed as internal controls. Full-length blots of each tested protein are reported in Supplementary Figure S5 . Error bars denote the mean ± S.D. of triplicate samples. * p

    Journal: Scientific Reports

    Article Title: PGC-1α attenuates hydrogen peroxide-induced apoptotic cell death by upregulating Nrf-2 via GSK3β inactivation mediated by activated p38 in HK-2 Cells

    doi: 10.1038/s41598-017-04593-w

    Figure Lengend Snippet: Upregulation of Nrf-2 by PGC-1α overexpression. The change in Nrf-2 mRNA and protein levels by PGC-1α overexpression was evaluated in stable cells. Cells were treated with 0.5 mM H 2 O 2 for 6 h. The mRNA levels of PGC-1α ( A ) and Nrf-2 ( B ) were analyzed by real-time PCR with each PCR primer pair. GAPDH was analyzed as an internal control. ( C ) Upregulation of Nrf-2 protein in cytosol and nucleus fraction by PGC-1α overexpression. ( D ) Regulation of Nrf-2 mediated HO-1 expression by PGC-1α. To assess the involvement between PGC-1α expression and Nrf-2 mediated HO-1 (as a one of well-known downstream molecules activated by Nrf-2) expression for protective effect of PGC-1α, PGC-1α stable cells were incubated with Nrf-2 specific siRNA (30 and 50 nM), and then, after 2 days, cells were treated with 0.5 mM H 2 O 2 for 6 h. The protein level of Nrf-2 and HO-1 were determined by immunoblotting with specific antibodies. Relative protein level was expressed as fold increases normalized to the untreated Mock cells. β-actin levels were analyzed as internal controls. Full-length blots of each tested protein are reported in Supplementary Figure S5 . Error bars denote the mean ± S.D. of triplicate samples. * p

    Article Snippet: For over-expression of human PGC-1α in HK-2 cells, human PGC-1α/pCDNA4 plasmid was purchased from Addgene (Cat#10974, Cambridge, USA).

    Techniques: Pyrolysis Gas Chromatography, Over Expression, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Expressing, Incubation

    Mutant huntingtin results in repressed transcriptional activities, and reduced PPARγ activity is independent of the protein level. B3 and E4 STHdh Q7 cells and 1A and 6L STHdh Q111 cells were transiently transfected with PPRE, CRE, or PGC-1α promoter luciferase reporter plasmids. The basal activities of PPRE ( A ), CRE ( B ), and PGC-1α promoter ( C ) reporters were dramatically reduced in both 1A and 6L STHdh Q111 cells. n = 3–4 Data shown are mean ± SE. D , PPARγ protein levels were variable in B3 and E4 STHdh Q7 cells and 1A and 6L STHdh Q111 cells, while the original STHdh Q111 cells exhibited lower levels of PPARγ than the original STHdh Q7 cells. Sixty micrograms of protein was run in each lane.

    Journal: PLoS ONE

    Article Title: Metabolic State Determines Sensitivity to Cellular Stress in Huntington Disease: Normalization by Activation of PPAR?

    doi: 10.1371/journal.pone.0030406

    Figure Lengend Snippet: Mutant huntingtin results in repressed transcriptional activities, and reduced PPARγ activity is independent of the protein level. B3 and E4 STHdh Q7 cells and 1A and 6L STHdh Q111 cells were transiently transfected with PPRE, CRE, or PGC-1α promoter luciferase reporter plasmids. The basal activities of PPRE ( A ), CRE ( B ), and PGC-1α promoter ( C ) reporters were dramatically reduced in both 1A and 6L STHdh Q111 cells. n = 3–4 Data shown are mean ± SE. D , PPARγ protein levels were variable in B3 and E4 STHdh Q7 cells and 1A and 6L STHdh Q111 cells, while the original STHdh Q111 cells exhibited lower levels of PPARγ than the original STHdh Q7 cells. Sixty micrograms of protein was run in each lane.

    Article Snippet: Plasmid Constructs PPRE×3-TK-Luc and human PGC-1α promoter-Luc were obtained from Addgene , .

    Techniques: Mutagenesis, Activity Assay, Transfection, Pyrolysis Gas Chromatography, Luciferase

    Schematic model of Erk5 protection against metabolic stress. FFA induces escalated ROS production from Gp91phox, which activates caplain-1, leading to a breakdown of Erk5. Loss of Erk5 and resultant Pgc-1α downregulation impose detrimental impacts on mitochondrial functions through accumulation of toxic lipids and vicious cycle of ROS. Prevention of Erk5 degradation or Erk5 restoration by AAV9 approach revives mitochondrial functions

    Journal: Nature Communications

    Article Title: Metabolic stress-induced cardiomyopathy is caused by mitochondrial dysfunction due to attenuated Erk5 signaling

    doi: 10.1038/s41467-017-00664-8

    Figure Lengend Snippet: Schematic model of Erk5 protection against metabolic stress. FFA induces escalated ROS production from Gp91phox, which activates caplain-1, leading to a breakdown of Erk5. Loss of Erk5 and resultant Pgc-1α downregulation impose detrimental impacts on mitochondrial functions through accumulation of toxic lipids and vicious cycle of ROS. Prevention of Erk5 degradation or Erk5 restoration by AAV9 approach revives mitochondrial functions

    Article Snippet: Adult rat cardiomyocytes transfection with siRNA or infection with adenovirus ARCMs were transfected with control siRNA, or rat Erk5 siRNA, rat calpain-1 siRNA (100 nM, siGENOME SMART pool, gene ID # 29153, Dharmacon), or rat Gp91phox siRNA (100 nM, # 66021) using Lipofectamine Plus reagent according to the manufacturer’s instruction (Invitrogen) for 48 h. Erk5-deficient ARCMs were then infected with adenovirus encoding human Pgc-1α (Vector Biolabs) at MOI 25 for 24 h prior to 2-hour PA treatment.

    Techniques: Pyrolysis Gas Chromatography

    Molecular basis underlying Erk5 regulation of Pgc-1α and Erk5 degradation. a Immunoblot analyses revealed the phosphorylation level and total expression of Erk5, Mef2a, Mef2c, Mef2d, and Pgc-1α in the hearts of Flox and CKO mice fed with chow or HFD for 16 weeks. b Immunoblot analyses showed increased activation of Erk5 in NRCMs treated with palmitate acid (PA, 500 µM) for 2 h; while the total expression of Erk5 is significantly decreased by PA for 8 h. c NRCMs were stained with DAPI ( blue ), anti-α-actinin antibody ( green ), and Mef2a ( red ) ( scale bar : 20 μm), fluorescence intensity of Mef2a in nuclei was quantified. d Increased Pgc-1α reporter luciferase activity was detected with overexpression of Erk5 or MEef2a. The increased reporter activity induced by palmitate was blunted by knockdown of Erk5 in NRCMs. e ChIP analysis demonstrated that enhanced binding of Mef2a to the Ppargca (Pgc-1α) promoter region at two sites (TSS, transcriptional starting site, -1506 and -2868, marked in red ) in HL-1 cardiomyocytes is Erk5-dependent. Sequence analysis of the purified PCR fragment bound by anti-Mef2a antibody confirmed the Mef2 consensus binding sequence. f Immunoblot analyses showed decreased Erk5 in ARCMs treated with 8 h PA or stearate acid (SA, 500 µM), but not by unsaturated FFA linoleate acid (LA, 500 µM) or oleate acid (OA, 500 µM). qPCR analysis showed no reduction in mRNA expression of Erk5 in PA or SA-treated ARCMs. g Immunoblot analyses showed that degradation of Erk5 was blocked by pretreatment of E-64D (25 μM, 4 h), but not by MG132 (10 μM, 4 h). Erk5 degradation was caused by calpain-1 and its expression could be restored by calpeptin (20 μM, 6 h), MDL-28170 (30 μM, 16 h) or Tempol (10 nM, 16 h), but not by cathepsin K inhibitor II (1 μM, 6 h). h Calpain-1 expression and its activity were measured. i Increased calpain activity, Nadph oxidase activity and DHE fluorescence intensity induced by PA were inhibited by pretreatment of apocynin (10 µM, 1 h) or gp91phox knockdown. Application of apocynin or Gp91phox knockdown restored Erk5 expression despite palmitate stimulation, n = 5–6 independent experiments per group. Actin is the protein loading control. Data are presented as means ± SD

    Journal: Nature Communications

    Article Title: Metabolic stress-induced cardiomyopathy is caused by mitochondrial dysfunction due to attenuated Erk5 signaling

    doi: 10.1038/s41467-017-00664-8

    Figure Lengend Snippet: Molecular basis underlying Erk5 regulation of Pgc-1α and Erk5 degradation. a Immunoblot analyses revealed the phosphorylation level and total expression of Erk5, Mef2a, Mef2c, Mef2d, and Pgc-1α in the hearts of Flox and CKO mice fed with chow or HFD for 16 weeks. b Immunoblot analyses showed increased activation of Erk5 in NRCMs treated with palmitate acid (PA, 500 µM) for 2 h; while the total expression of Erk5 is significantly decreased by PA for 8 h. c NRCMs were stained with DAPI ( blue ), anti-α-actinin antibody ( green ), and Mef2a ( red ) ( scale bar : 20 μm), fluorescence intensity of Mef2a in nuclei was quantified. d Increased Pgc-1α reporter luciferase activity was detected with overexpression of Erk5 or MEef2a. The increased reporter activity induced by palmitate was blunted by knockdown of Erk5 in NRCMs. e ChIP analysis demonstrated that enhanced binding of Mef2a to the Ppargca (Pgc-1α) promoter region at two sites (TSS, transcriptional starting site, -1506 and -2868, marked in red ) in HL-1 cardiomyocytes is Erk5-dependent. Sequence analysis of the purified PCR fragment bound by anti-Mef2a antibody confirmed the Mef2 consensus binding sequence. f Immunoblot analyses showed decreased Erk5 in ARCMs treated with 8 h PA or stearate acid (SA, 500 µM), but not by unsaturated FFA linoleate acid (LA, 500 µM) or oleate acid (OA, 500 µM). qPCR analysis showed no reduction in mRNA expression of Erk5 in PA or SA-treated ARCMs. g Immunoblot analyses showed that degradation of Erk5 was blocked by pretreatment of E-64D (25 μM, 4 h), but not by MG132 (10 μM, 4 h). Erk5 degradation was caused by calpain-1 and its expression could be restored by calpeptin (20 μM, 6 h), MDL-28170 (30 μM, 16 h) or Tempol (10 nM, 16 h), but not by cathepsin K inhibitor II (1 μM, 6 h). h Calpain-1 expression and its activity were measured. i Increased calpain activity, Nadph oxidase activity and DHE fluorescence intensity induced by PA were inhibited by pretreatment of apocynin (10 µM, 1 h) or gp91phox knockdown. Application of apocynin or Gp91phox knockdown restored Erk5 expression despite palmitate stimulation, n = 5–6 independent experiments per group. Actin is the protein loading control. Data are presented as means ± SD

    Article Snippet: Adult rat cardiomyocytes transfection with siRNA or infection with adenovirus ARCMs were transfected with control siRNA, or rat Erk5 siRNA, rat calpain-1 siRNA (100 nM, siGENOME SMART pool, gene ID # 29153, Dharmacon), or rat Gp91phox siRNA (100 nM, # 66021) using Lipofectamine Plus reagent according to the manufacturer’s instruction (Invitrogen) for 48 h. Erk5-deficient ARCMs were then infected with adenovirus encoding human Pgc-1α (Vector Biolabs) at MOI 25 for 24 h prior to 2-hour PA treatment.

    Techniques: Pyrolysis Gas Chromatography, Expressing, Mouse Assay, Activation Assay, Staining, Fluorescence, Luciferase, Activity Assay, Over Expression, Chromatin Immunoprecipitation, Binding Assay, Sequencing, Purification, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    Identification of LDLR promoter region that is responsible for PGC-1α-mediated suppression. The original pLR1563-luc and the indicated series of 5'-deletion constructs linked to luciferase were cotransfected into HepG2 cells with LacZ expression

    Journal:

    Article Title: Sterol-independent repression of low density lipoprotein receptor promoter by peroxisome proliferator activated receptor ? coactivator-1? (PGC-1?)

    doi: 10.3858/emm.2009.41.6.046

    Figure Lengend Snippet: Identification of LDLR promoter region that is responsible for PGC-1α-mediated suppression. The original pLR1563-luc and the indicated series of 5'-deletion constructs linked to luciferase were cotransfected into HepG2 cells with LacZ expression

    Article Snippet: Protein extracts (20 µg) from HepG2 cells infected with AdLacZ or AdPGC-1 adenovirus were prepared with lysis buffer (10 mM HEPES, pH 7.9, 10 mM KCl, 2 mM MgCl2 , 0.5 mM dithiothreitol, 1 mM PMSF, 5 µg/ml aprotinin, 5 µg/ml pepstatin A, 5 µg/ml leupeptin, and 1% Triton X-100), separated by 10% SDS-PAGE, and analyzed by Western blot using rabbit anti-human PGC-1α polyclonal antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) and an enhanced chemiluminescence system (ECL, Amersham Pharmacia Biotech).

    Techniques: Pyrolysis Gas Chromatography, Construct, Luciferase, Expressing

    PGC-1α may interact with the LDLR distal promoter through unknown factor. (A) The distal LDLR promoter (-974 to -633) contains E-box and AP1 binding motifs. (B) ChIP assay. HepG2 cells were infected with Ad-PGC-1 or Ad-LacZ control. After 36 h,

    Journal:

    Article Title: Sterol-independent repression of low density lipoprotein receptor promoter by peroxisome proliferator activated receptor ? coactivator-1? (PGC-1?)

    doi: 10.3858/emm.2009.41.6.046

    Figure Lengend Snippet: PGC-1α may interact with the LDLR distal promoter through unknown factor. (A) The distal LDLR promoter (-974 to -633) contains E-box and AP1 binding motifs. (B) ChIP assay. HepG2 cells were infected with Ad-PGC-1 or Ad-LacZ control. After 36 h,

    Article Snippet: Protein extracts (20 µg) from HepG2 cells infected with AdLacZ or AdPGC-1 adenovirus were prepared with lysis buffer (10 mM HEPES, pH 7.9, 10 mM KCl, 2 mM MgCl2 , 0.5 mM dithiothreitol, 1 mM PMSF, 5 µg/ml aprotinin, 5 µg/ml pepstatin A, 5 µg/ml leupeptin, and 1% Triton X-100), separated by 10% SDS-PAGE, and analyzed by Western blot using rabbit anti-human PGC-1α polyclonal antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) and an enhanced chemiluminescence system (ECL, Amersham Pharmacia Biotech).

    Techniques: Pyrolysis Gas Chromatography, Binding Assay, Chromatin Immunoprecipitation, Infection

    Inhibition of p38-MAPK had no effect on the PGC-1α-repressed LDLR mRNA expression. HepG2 cells were infected with AdLacZ alone (AdPGC -) or AdPGC-1 (AdPGC +) for 24 h at a multiplicity of infection of 50. The infected cells in serum free media

    Journal:

    Article Title: Sterol-independent repression of low density lipoprotein receptor promoter by peroxisome proliferator activated receptor ? coactivator-1? (PGC-1?)

    doi: 10.3858/emm.2009.41.6.046

    Figure Lengend Snippet: Inhibition of p38-MAPK had no effect on the PGC-1α-repressed LDLR mRNA expression. HepG2 cells were infected with AdLacZ alone (AdPGC -) or AdPGC-1 (AdPGC +) for 24 h at a multiplicity of infection of 50. The infected cells in serum free media

    Article Snippet: Protein extracts (20 µg) from HepG2 cells infected with AdLacZ or AdPGC-1 adenovirus were prepared with lysis buffer (10 mM HEPES, pH 7.9, 10 mM KCl, 2 mM MgCl2 , 0.5 mM dithiothreitol, 1 mM PMSF, 5 µg/ml aprotinin, 5 µg/ml pepstatin A, 5 µg/ml leupeptin, and 1% Triton X-100), separated by 10% SDS-PAGE, and analyzed by Western blot using rabbit anti-human PGC-1α polyclonal antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) and an enhanced chemiluminescence system (ECL, Amersham Pharmacia Biotech).

    Techniques: Inhibition, Pyrolysis Gas Chromatography, Expressing, Infection

    Repression of LDLR promoter activity by PGC-1α that co-activates the ERα/ERE-dependent transcription. HepG2 cells were transfected with LacZ expression vector (400 ng), expression vectors (400 ng) for PGC-1α and/or ERα,

    Journal:

    Article Title: Sterol-independent repression of low density lipoprotein receptor promoter by peroxisome proliferator activated receptor ? coactivator-1? (PGC-1?)

    doi: 10.3858/emm.2009.41.6.046

    Figure Lengend Snippet: Repression of LDLR promoter activity by PGC-1α that co-activates the ERα/ERE-dependent transcription. HepG2 cells were transfected with LacZ expression vector (400 ng), expression vectors (400 ng) for PGC-1α and/or ERα,

    Article Snippet: Protein extracts (20 µg) from HepG2 cells infected with AdLacZ or AdPGC-1 adenovirus were prepared with lysis buffer (10 mM HEPES, pH 7.9, 10 mM KCl, 2 mM MgCl2 , 0.5 mM dithiothreitol, 1 mM PMSF, 5 µg/ml aprotinin, 5 µg/ml pepstatin A, 5 µg/ml leupeptin, and 1% Triton X-100), separated by 10% SDS-PAGE, and analyzed by Western blot using rabbit anti-human PGC-1α polyclonal antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) and an enhanced chemiluminescence system (ECL, Amersham Pharmacia Biotech).

    Techniques: Activity Assay, Pyrolysis Gas Chromatography, Transfection, Expressing, Plasmid Preparation

    PGC-1α repressed the LDLR promoter activity regardless of cholesterol. HepG2 cells were transfected with LacZ expression vector (400 ng), expression vectors (400 ng) for PGC-1α or mock vector (pcDNA3.1/HisC), and pLR1563-luc, as indicated.

    Journal:

    Article Title: Sterol-independent repression of low density lipoprotein receptor promoter by peroxisome proliferator activated receptor ? coactivator-1? (PGC-1?)

    doi: 10.3858/emm.2009.41.6.046

    Figure Lengend Snippet: PGC-1α repressed the LDLR promoter activity regardless of cholesterol. HepG2 cells were transfected with LacZ expression vector (400 ng), expression vectors (400 ng) for PGC-1α or mock vector (pcDNA3.1/HisC), and pLR1563-luc, as indicated.

    Article Snippet: Protein extracts (20 µg) from HepG2 cells infected with AdLacZ or AdPGC-1 adenovirus were prepared with lysis buffer (10 mM HEPES, pH 7.9, 10 mM KCl, 2 mM MgCl2 , 0.5 mM dithiothreitol, 1 mM PMSF, 5 µg/ml aprotinin, 5 µg/ml pepstatin A, 5 µg/ml leupeptin, and 1% Triton X-100), separated by 10% SDS-PAGE, and analyzed by Western blot using rabbit anti-human PGC-1α polyclonal antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) and an enhanced chemiluminescence system (ECL, Amersham Pharmacia Biotech).

    Techniques: Pyrolysis Gas Chromatography, Activity Assay, Transfection, Expressing, Plasmid Preparation

    SRE-1 is not required for PGC-1α-mediated inhibition of LDLR promoter activity. HepG2 cells were transfected with LacZ expression vector, expression vectors (400 ng) for PGC-1α or mock vector (pcDNA3.1/HisC), and one of three LDLR promoter

    Journal:

    Article Title: Sterol-independent repression of low density lipoprotein receptor promoter by peroxisome proliferator activated receptor ? coactivator-1? (PGC-1?)

    doi: 10.3858/emm.2009.41.6.046

    Figure Lengend Snippet: SRE-1 is not required for PGC-1α-mediated inhibition of LDLR promoter activity. HepG2 cells were transfected with LacZ expression vector, expression vectors (400 ng) for PGC-1α or mock vector (pcDNA3.1/HisC), and one of three LDLR promoter

    Article Snippet: Protein extracts (20 µg) from HepG2 cells infected with AdLacZ or AdPGC-1 adenovirus were prepared with lysis buffer (10 mM HEPES, pH 7.9, 10 mM KCl, 2 mM MgCl2 , 0.5 mM dithiothreitol, 1 mM PMSF, 5 µg/ml aprotinin, 5 µg/ml pepstatin A, 5 µg/ml leupeptin, and 1% Triton X-100), separated by 10% SDS-PAGE, and analyzed by Western blot using rabbit anti-human PGC-1α polyclonal antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) and an enhanced chemiluminescence system (ECL, Amersham Pharmacia Biotech).

    Techniques: Pyrolysis Gas Chromatography, Inhibition, Activity Assay, Transfection, Expressing, Plasmid Preparation

    Effect of N-SREBP-2 dominant positive or PPARγ activation on the PGC-1α-repressed LDLR promoter activity. (A) Overexpression of N-SREBP2 dominates the repressive effect of PGC-1α. HepG2 cells were transfected with LacZ expression

    Journal:

    Article Title: Sterol-independent repression of low density lipoprotein receptor promoter by peroxisome proliferator activated receptor ? coactivator-1? (PGC-1?)

    doi: 10.3858/emm.2009.41.6.046

    Figure Lengend Snippet: Effect of N-SREBP-2 dominant positive or PPARγ activation on the PGC-1α-repressed LDLR promoter activity. (A) Overexpression of N-SREBP2 dominates the repressive effect of PGC-1α. HepG2 cells were transfected with LacZ expression

    Article Snippet: Protein extracts (20 µg) from HepG2 cells infected with AdLacZ or AdPGC-1 adenovirus were prepared with lysis buffer (10 mM HEPES, pH 7.9, 10 mM KCl, 2 mM MgCl2 , 0.5 mM dithiothreitol, 1 mM PMSF, 5 µg/ml aprotinin, 5 µg/ml pepstatin A, 5 µg/ml leupeptin, and 1% Triton X-100), separated by 10% SDS-PAGE, and analyzed by Western blot using rabbit anti-human PGC-1α polyclonal antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) and an enhanced chemiluminescence system (ECL, Amersham Pharmacia Biotech).

    Techniques: Activation Assay, Pyrolysis Gas Chromatography, Activity Assay, Over Expression, Transfection, Expressing

    Repression of endogenous LDLR mRNA by overexpression of PGC-1α. HepG2 cells were infected with AdLacZ or AdPGC-1α for 48 h at a multiplicity of infection of 50. Total RNA or cell lysate was prepared from the cells. RT-PCR (RT), Northern

    Journal:

    Article Title: Sterol-independent repression of low density lipoprotein receptor promoter by peroxisome proliferator activated receptor ? coactivator-1? (PGC-1?)

    doi: 10.3858/emm.2009.41.6.046

    Figure Lengend Snippet: Repression of endogenous LDLR mRNA by overexpression of PGC-1α. HepG2 cells were infected with AdLacZ or AdPGC-1α for 48 h at a multiplicity of infection of 50. Total RNA or cell lysate was prepared from the cells. RT-PCR (RT), Northern

    Article Snippet: Protein extracts (20 µg) from HepG2 cells infected with AdLacZ or AdPGC-1 adenovirus were prepared with lysis buffer (10 mM HEPES, pH 7.9, 10 mM KCl, 2 mM MgCl2 , 0.5 mM dithiothreitol, 1 mM PMSF, 5 µg/ml aprotinin, 5 µg/ml pepstatin A, 5 µg/ml leupeptin, and 1% Triton X-100), separated by 10% SDS-PAGE, and analyzed by Western blot using rabbit anti-human PGC-1α polyclonal antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) and an enhanced chemiluminescence system (ECL, Amersham Pharmacia Biotech).

    Techniques: Over Expression, Pyrolysis Gas Chromatography, Infection, Reverse Transcription Polymerase Chain Reaction, Northern Blot

    Changes in VEGF protein levels Effect of PGC-1α siRNA on VEGF protein production in hRVECs at 16h of normoxia and hypoxia after transfection were detected by ELISA. The VEGF protein production in siPGC-1α groups were significantly downregulated when compared with control siRNA groups under normoxia and hypoxia. b P

    Journal: International Journal of Ophthalmology

    Article Title: Small interfering RNA targeting PGC-1α inhibits VEGF expression and tube formation in human retinal vascular endothelial cells

    doi: 10.3980/j.issn.2222-3959.2015.05.05

    Figure Lengend Snippet: Changes in VEGF protein levels Effect of PGC-1α siRNA on VEGF protein production in hRVECs at 16h of normoxia and hypoxia after transfection were detected by ELISA. The VEGF protein production in siPGC-1α groups were significantly downregulated when compared with control siRNA groups under normoxia and hypoxia. b P

    Article Snippet: The primary antibodies used were as follows: mouse anti-human PGC-1α (Abcam, Cambridge, MA, USA) 1:1000, mouse anti-human β-actin (Santa Cruz Biotechnology, Dallas, TX, USA) 1:800.

    Techniques: Pyrolysis Gas Chromatography, Transfection, Enzyme-linked Immunosorbent Assay

    Changes in PGC-1α mRNA expression PGC-1α siRNA-mediated downregulation of PGC-1α mRNA expression in hRVECs at 16h of normoxia and hypoxia after transfection were examined by real-time PCR. The expression of PGC-1α mRNA were significantly downregulated in cells in the siPGC-1α groups relative to cells in the control siRNA groups under normoxia and hypoxia. b P

    Journal: International Journal of Ophthalmology

    Article Title: Small interfering RNA targeting PGC-1α inhibits VEGF expression and tube formation in human retinal vascular endothelial cells

    doi: 10.3980/j.issn.2222-3959.2015.05.05

    Figure Lengend Snippet: Changes in PGC-1α mRNA expression PGC-1α siRNA-mediated downregulation of PGC-1α mRNA expression in hRVECs at 16h of normoxia and hypoxia after transfection were examined by real-time PCR. The expression of PGC-1α mRNA were significantly downregulated in cells in the siPGC-1α groups relative to cells in the control siRNA groups under normoxia and hypoxia. b P

    Article Snippet: The primary antibodies used were as follows: mouse anti-human PGC-1α (Abcam, Cambridge, MA, USA) 1:1000, mouse anti-human β-actin (Santa Cruz Biotechnology, Dallas, TX, USA) 1:800.

    Techniques: Pyrolysis Gas Chromatography, Expressing, Transfection, Real-time Polymerase Chain Reaction

    PGC-1α siRNA inhibits cell tube formation in hRVECs Tubular structures were photographed (A-F) and then quantified by counting the number of branch points (G) and calculating total tube length (H) in cultured hRVECs. A: Normoxia siPGC-1α group; B: Normoxia control siRNA group; C: Normoxia without siRNA group; D: Hypoxia siPGC-1α group; E: Hypoxia control siRNA group; F: Hypoxia without siRNA group. PGC-1α siRNA significantly reduced tubular-like formation (A and D), branch points (G), and total tube length (H) in cells compared with control siRNA group under normoxia (B) and hypoxia (E). Cells in the hypoxia without siRNA group and the hypoxia control siRNA group formed significantly more tubes than cells in their corresponding normoxia groups. b P

    Journal: International Journal of Ophthalmology

    Article Title: Small interfering RNA targeting PGC-1α inhibits VEGF expression and tube formation in human retinal vascular endothelial cells

    doi: 10.3980/j.issn.2222-3959.2015.05.05

    Figure Lengend Snippet: PGC-1α siRNA inhibits cell tube formation in hRVECs Tubular structures were photographed (A-F) and then quantified by counting the number of branch points (G) and calculating total tube length (H) in cultured hRVECs. A: Normoxia siPGC-1α group; B: Normoxia control siRNA group; C: Normoxia without siRNA group; D: Hypoxia siPGC-1α group; E: Hypoxia control siRNA group; F: Hypoxia without siRNA group. PGC-1α siRNA significantly reduced tubular-like formation (A and D), branch points (G), and total tube length (H) in cells compared with control siRNA group under normoxia (B) and hypoxia (E). Cells in the hypoxia without siRNA group and the hypoxia control siRNA group formed significantly more tubes than cells in their corresponding normoxia groups. b P

    Article Snippet: The primary antibodies used were as follows: mouse anti-human PGC-1α (Abcam, Cambridge, MA, USA) 1:1000, mouse anti-human β-actin (Santa Cruz Biotechnology, Dallas, TX, USA) 1:800.

    Techniques: Pyrolysis Gas Chromatography, Cell Culture

    Changes in PGC-1α protein levels A: PGC-1α siRNA-mediated downregulation of PGC-1α protein production in hRVECs at 16h of normoxia and hypoxia after transfection were detected by Western blot; B: Quantitative results of the Western blot analysis in A.The levels of PGC-1α protein were significantly lower in cells in the siPGC-1α groups compared with cells in the control siRNA groups under normoxia and hypoxia. b P

    Journal: International Journal of Ophthalmology

    Article Title: Small interfering RNA targeting PGC-1α inhibits VEGF expression and tube formation in human retinal vascular endothelial cells

    doi: 10.3980/j.issn.2222-3959.2015.05.05

    Figure Lengend Snippet: Changes in PGC-1α protein levels A: PGC-1α siRNA-mediated downregulation of PGC-1α protein production in hRVECs at 16h of normoxia and hypoxia after transfection were detected by Western blot; B: Quantitative results of the Western blot analysis in A.The levels of PGC-1α protein were significantly lower in cells in the siPGC-1α groups compared with cells in the control siRNA groups under normoxia and hypoxia. b P

    Article Snippet: The primary antibodies used were as follows: mouse anti-human PGC-1α (Abcam, Cambridge, MA, USA) 1:1000, mouse anti-human β-actin (Santa Cruz Biotechnology, Dallas, TX, USA) 1:800.

    Techniques: Pyrolysis Gas Chromatography, Transfection, Western Blot

    Changes in VEGF mRNA expression Effect of PGC-1α siRNA on VEGF mRNA expression in hRVECs at 16h of normoxia and hypoxia after transfection were examined by real-time PCR. The VEGF mRNA levels of siPGC-1α groups were significantly downregulated when compared with control siRNA groups during normoxia and hypoxia. b P

    Journal: International Journal of Ophthalmology

    Article Title: Small interfering RNA targeting PGC-1α inhibits VEGF expression and tube formation in human retinal vascular endothelial cells

    doi: 10.3980/j.issn.2222-3959.2015.05.05

    Figure Lengend Snippet: Changes in VEGF mRNA expression Effect of PGC-1α siRNA on VEGF mRNA expression in hRVECs at 16h of normoxia and hypoxia after transfection were examined by real-time PCR. The VEGF mRNA levels of siPGC-1α groups were significantly downregulated when compared with control siRNA groups during normoxia and hypoxia. b P

    Article Snippet: The primary antibodies used were as follows: mouse anti-human PGC-1α (Abcam, Cambridge, MA, USA) 1:1000, mouse anti-human β-actin (Santa Cruz Biotechnology, Dallas, TX, USA) 1:800.

    Techniques: Expressing, Pyrolysis Gas Chromatography, Transfection, Real-time Polymerase Chain Reaction

    PGC-1α siRNA inhibits cell proliferation in hRVECs Effect of PGC-1α siRNA on cell proliferation in hRVECs at 16h of normoxia and hypoxia after transfection. The percentage of BrdU-labeled cells in siPGC-1α groups significantly decreased compared with control siRNA groups under normoxia and hypoxia. b P

    Journal: International Journal of Ophthalmology

    Article Title: Small interfering RNA targeting PGC-1α inhibits VEGF expression and tube formation in human retinal vascular endothelial cells

    doi: 10.3980/j.issn.2222-3959.2015.05.05

    Figure Lengend Snippet: PGC-1α siRNA inhibits cell proliferation in hRVECs Effect of PGC-1α siRNA on cell proliferation in hRVECs at 16h of normoxia and hypoxia after transfection. The percentage of BrdU-labeled cells in siPGC-1α groups significantly decreased compared with control siRNA groups under normoxia and hypoxia. b P

    Article Snippet: The primary antibodies used were as follows: mouse anti-human PGC-1α (Abcam, Cambridge, MA, USA) 1:1000, mouse anti-human β-actin (Santa Cruz Biotechnology, Dallas, TX, USA) 1:800.

    Techniques: Pyrolysis Gas Chromatography, Transfection, Labeling

    Endothelial PGC-1α overexpression protects from endothelial dysfunction. ( A ) PGC-1α endothelial specific transgene construct with representative expression of human (h) and mouse (m) PGC-1α mRNA in MLECs from the indicated genotype. ( B ) MLEC mRNA quantification (N = 3, * P

    Journal: Scientific Reports

    Article Title: PGC-1α dictates endothelial function through regulation of eNOS expression

    doi: 10.1038/srep38210

    Figure Lengend Snippet: Endothelial PGC-1α overexpression protects from endothelial dysfunction. ( A ) PGC-1α endothelial specific transgene construct with representative expression of human (h) and mouse (m) PGC-1α mRNA in MLECs from the indicated genotype. ( B ) MLEC mRNA quantification (N = 3, * P

    Article Snippet: Generation of endothelial specific PGC-1α manipulation in mice Human PGC-1α cDNA (Origene, Rockville, MD) was linearized and inserted into pBSmVELacZ (Obtained from Kenneth Walsh, Ph.D. Boston University, Boston, MA) to replace the LacZ open reading frame through the NotI site to produce human PGC-1α expression under the control of the mouse vascular endothelial cadherin promoter (VE-Cad; ).

    Techniques: Pyrolysis Gas Chromatography, Over Expression, Construct, Expressing

    Loss of endothelial PGC-1α leads to reduced NO• bioactivity. ( A ) WT mice received a pressor dose of Angiotensin II (ATII; 1 mg/kg; 7 days). Mouse lung endothelial cells (MLECs) were harvested and PGC-1α mRNA expression measured (N = 4/group, * P

    Journal: Scientific Reports

    Article Title: PGC-1α dictates endothelial function through regulation of eNOS expression

    doi: 10.1038/srep38210

    Figure Lengend Snippet: Loss of endothelial PGC-1α leads to reduced NO• bioactivity. ( A ) WT mice received a pressor dose of Angiotensin II (ATII; 1 mg/kg; 7 days). Mouse lung endothelial cells (MLECs) were harvested and PGC-1α mRNA expression measured (N = 4/group, * P

    Article Snippet: Generation of endothelial specific PGC-1α manipulation in mice Human PGC-1α cDNA (Origene, Rockville, MD) was linearized and inserted into pBSmVELacZ (Obtained from Kenneth Walsh, Ph.D. Boston University, Boston, MA) to replace the LacZ open reading frame through the NotI site to produce human PGC-1α expression under the control of the mouse vascular endothelial cadherin promoter (VE-Cad; ).

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Expressing

    PGC-1α upregulates eNOS expression and limits hypertension-induced endothelial dysfunction. Human aortic endothelial cells (HAECs) were transfected or not with adenoviral (Ad) vectors containing β-galactosidase (Ctl) or PGC-1α for 24 h and assessed for ( A ) cGMP (N = 3), eNOS ( B ) mRNA (N = 3) and ( C , D ) protein; * P

    Journal: Scientific Reports

    Article Title: PGC-1α dictates endothelial function through regulation of eNOS expression

    doi: 10.1038/srep38210

    Figure Lengend Snippet: PGC-1α upregulates eNOS expression and limits hypertension-induced endothelial dysfunction. Human aortic endothelial cells (HAECs) were transfected or not with adenoviral (Ad) vectors containing β-galactosidase (Ctl) or PGC-1α for 24 h and assessed for ( A ) cGMP (N = 3), eNOS ( B ) mRNA (N = 3) and ( C , D ) protein; * P

    Article Snippet: Generation of endothelial specific PGC-1α manipulation in mice Human PGC-1α cDNA (Origene, Rockville, MD) was linearized and inserted into pBSmVELacZ (Obtained from Kenneth Walsh, Ph.D. Boston University, Boston, MA) to replace the LacZ open reading frame through the NotI site to produce human PGC-1α expression under the control of the mouse vascular endothelial cadherin promoter (VE-Cad; ).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Transfection, CTL Assay

    ERRα is required for PGC-1α dependent eNOS expression. ( A ) HAECs were transfected with adenoviral vectors expressing β-galactosidase (Ad-Ctl) or PGC-1α (Ad- PGC-1α) and mRNA expression of the indicated genes determined by RT-PCR (N = 3, *P

    Journal: Scientific Reports

    Article Title: PGC-1α dictates endothelial function through regulation of eNOS expression

    doi: 10.1038/srep38210

    Figure Lengend Snippet: ERRα is required for PGC-1α dependent eNOS expression. ( A ) HAECs were transfected with adenoviral vectors expressing β-galactosidase (Ad-Ctl) or PGC-1α (Ad- PGC-1α) and mRNA expression of the indicated genes determined by RT-PCR (N = 3, *P

    Article Snippet: Generation of endothelial specific PGC-1α manipulation in mice Human PGC-1α cDNA (Origene, Rockville, MD) was linearized and inserted into pBSmVELacZ (Obtained from Kenneth Walsh, Ph.D. Boston University, Boston, MA) to replace the LacZ open reading frame through the NotI site to produce human PGC-1α expression under the control of the mouse vascular endothelial cadherin promoter (VE-Cad; ).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Transfection, CTL Assay, Reverse Transcription Polymerase Chain Reaction

    Plasmid construct for generation of actin-PGC-1α transgenic mice. A Not I site was inserted at the vector Xba I site and a Sal I site inserted at the vector Hind III site on the pCAGGS plasmid. Human peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC-1α) cDNA was inserted in the Not I site and Sal I digestion released the fragment containing the cytomegalovirus immediate-early (CMV-IE) enhancer, chicken β-actin promoter, and hPGC-1α that was used for microinjection.

    Journal: Biomolecules

    Article Title: Peroxisome Proliferator Activator Receptor Gamma Coactivator-1α Overexpression in Amyotrophic Lateral Sclerosis: A Tale of Two Transgenics

    doi: 10.3390/biom10050760

    Figure Lengend Snippet: Plasmid construct for generation of actin-PGC-1α transgenic mice. A Not I site was inserted at the vector Xba I site and a Sal I site inserted at the vector Hind III site on the pCAGGS plasmid. Human peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC-1α) cDNA was inserted in the Not I site and Sal I digestion released the fragment containing the cytomegalovirus immediate-early (CMV-IE) enhancer, chicken β-actin promoter, and hPGC-1α that was used for microinjection.

    Article Snippet: Then, the 3.1 kb human PGC-1α (hPGC-1α) cDNA (NM_013261.2; obtained from OriGene Technologies, Inc., Rockville, MD, USA) was inserted in the Not I site , followed by SalI digestion to release a ~6 kb fragment containing the CMV-IE enhancer, chicken β-actin promoter, and hPGC-1α.

    Techniques: Plasmid Preparation, Construct, Pyrolysis Gas Chromatography, Transgenic Assay, Mouse Assay