pgc-1α Search Results


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  • 98
    Millipore pgc 1α
    PolG mice display higher levels of mitochondrial regulatory proteins and transcription factors. AMPKα activation ( A ), <t>PGC-1α</t> ( B ), NRF-1 ( C ), and Tfam ( D ) were determined by Western Blotting in muscle from young (3–6 mo) and older (8–15 mo) WT and PolG animals. AMPKα activation is determined by phosphorylated AMPKα over total AMPKα. Representative blots are shown above with a graphical summary of the data below (n = 7–13). Significance was set at P
    Pgc 1α, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 899 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgc 1α/product/Millipore
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    94
    Santa Cruz Biotechnology pgc 1α
    AMPK activation on the expressions of AT 1 R and SIRT1, mitochondrial biogenesis and ROS level in RVLM of young offspring exposed to maternal HFD or ND. Representative gels and densitometric analysis of results from Western blot showing changes in protein expression of ( a ) AT 1 R; ( b ) SIRT1, ( c ) <t>PGC-1α,</t> or ( d ) TFAM; ( e ) mitochondrial DNA copy number; and ( f ) ROS level in RVLM of ND or HFD offspring, alone or with additional treatment with an AMPK activator, metformin (400 mg·kg − 1 ·day − 1 ), administered via gastric gavage at age of 8 weeks for 4 weeks. Analysis was performed on tissues collected bilaterally from individual RVLM at age of 12 weeks. Data on protein expression were normalized to the average ND control value, which is set to 1.0, and are presented as mean ± SEM ( n = 8 in all groups). * P
    Pgc 1α, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1565 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam pgc 1α
    <t>PGC-1α,</t> PPARα and GyK expression increases in KO mice. EATs were dissected from miR-150 KO (n = 7) and WT (n = 7) mice after 17 weeks HFD feeding. A) PGC-1α, PPARα and GyK gene expression was assessed by qRT-PCR. B) Protein levels of PGC-1α and GyK were measured by Western blotting. C) Western blotting results were quantified by ImageJ. PGC-1α and GyK expression were normalized by β-actin and GAPDH respectively. Data represent mean ± SEM, *p
    Pgc 1α, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgc 1α/product/Abcam
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    93
    Cell Signaling Technology Inc pgc 1α
    Effects of flavan-3-ols on expression of mitochondrial biogenesis-related protein in insulin-resistant 3T3-L1 adipocytes. RT-PCR analysis of mRNA levels of, ( a ) <t>PGC-1α,</t> ( b ) NRF1, and ( c ) Tfam. ( d ) Western blot analysis of expression of SirT1 and PGC-1α. Fully differentiated 3T3-L1 adipocytes were treated with Dex for 48 h in the presence or absence of flavan-3-ols or rosiglitazone. Con, normal differentiated 3T3-L1 adipocytes; Mod, Dex alone treated differentiated 3T3-L1 adipocytes; Rosi, rosiglitazone; C, catechin; EC, epicatechin; PB7, procyanidin B7; PB6, procyanidin B6; PB3, procyanidin B3; PB1, procyanidin B1. Values are shown as Mean ± SD of three independent triplicate experiments ( n = 3). # p
    Pgc 1α, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 443 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore anti pgc 1α
    Mitochondrial ultrastructural abnormalities in hearts of adult <t>PGC-1αβ</t> −/− mice. Representative electron micrographs of sections taken from the left ventricular papillary muscle from 16-week-old <t>PGC-1α</t> −/− (α −/− ) and PGC-1αβ −/− (αβ −/− ) mice at two different magnifications. Arrows indicate the structurally abnormal mitochondria with “stacked” cristae ( white arrows ). Scale bar for each magnification is shown at the bottom right corner : A , 1 μm; B , 5 μm.
    Anti Pgc 1α, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 222 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Novus Biologicals pgc1 alpha antibody
    Mitochondrial ultrastructural abnormalities in hearts of adult <t>PGC-1αβ</t> −/− mice. Representative electron micrographs of sections taken from the left ventricular papillary muscle from 16-week-old <t>PGC-1α</t> −/− (α −/− ) and PGC-1αβ −/− (αβ −/− ) mice at two different magnifications. Arrows indicate the structurally abnormal mitochondria with “stacked” cristae ( white arrows ). Scale bar for each magnification is shown at the bottom right corner : A , 1 μm; B , 5 μm.
    Pgc1 Alpha Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 98/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    PolG mice display higher levels of mitochondrial regulatory proteins and transcription factors. AMPKα activation ( A ), PGC-1α ( B ), NRF-1 ( C ), and Tfam ( D ) were determined by Western Blotting in muscle from young (3–6 mo) and older (8–15 mo) WT and PolG animals. AMPKα activation is determined by phosphorylated AMPKα over total AMPKα. Representative blots are shown above with a graphical summary of the data below (n = 7–13). Significance was set at P

    Journal: PLoS ONE

    Article Title: Dysregulation of Mitochondrial Quality Control Processes Contribute to Sarcopenia in a Mouse Model of Premature Aging

    doi: 10.1371/journal.pone.0069327

    Figure Lengend Snippet: PolG mice display higher levels of mitochondrial regulatory proteins and transcription factors. AMPKα activation ( A ), PGC-1α ( B ), NRF-1 ( C ), and Tfam ( D ) were determined by Western Blotting in muscle from young (3–6 mo) and older (8–15 mo) WT and PolG animals. AMPKα activation is determined by phosphorylated AMPKα over total AMPKα. Representative blots are shown above with a graphical summary of the data below (n = 7–13). Significance was set at P

    Article Snippet: After transfer, membranes were blocked by incubating for 1 hr at room temperature in Starting Block T20 Blocking Buffer (Thermo Scientific) and incubated overnight at 4°C in blocking buffer at a dilution of 1∶500 for Phosphorylated AMPKα and Total AMPKα (Cell Signaling, 2531 and 2532, respectively), 1∶500 for PGC-1α (Calbiochem, 516557), 1∶500 for NRF-1 (Santa Cruz, Sc-19050), 1∶1000 for Tfam (Calbiochem, DR-1071),1∶500 for Mfn1 (Santa Cruz, Sc-50330) and Mfn2 (Sigma, M6444), 1∶1000 for Opa1 and DRP1 (BD Transduction Laboratories, 612606 and 611112), 1∶500 for Fis1 (Alexis Biochemicals, ALX-210-907-R100), 1∶1000 for mtHsp70 and cHsp70 (Assay Designs, ADI-SPS-825 and ADI-SPS-810), 1∶500 for Tom22 (Sigma, T6319), 1∶1000 for Beclin-1 (Cell Signaling, 3738), 1∶1000 for LC3 (Cell Signaling, 2775), 1∶1000 for p62 (Sigma, P0067), 1∶500 for Atg5 (Cell Signaling, 2630), 1∶500 for ULK1 (Cell Signaling, 8054), 1∶5000 for actin (Sigma, A2066).

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

    Gene and protein expression in the EDL muscle (A–C). (A) Relative expression levels of genes for energy metabolism (myoglobin, LPL, UCP3, PDK4), mitochondrial proteins (porin, MTCO2, UCP3 and PDK4), and transcription factors (PPARδ, FOXO1, MyoD, myogenin) and a coactivator (PGC1α) for muscle fiber type regulation in EDL muscles from soybean oil-fed (open bars), fish oil-fed (filled bars), and lard-fed (gray bars) rats. Each gene was normalized to β-actin mRNA. Values are expressed as fold changes compared with the soybean oil-fed group. (B) Protein expression levels of fiber type-related markers (myoglobin, UCP3, porin, PGC1α, and FOXO1) and loading control (actin) in the EDL, with densitometry quantification in panel C. The two arrowheads in porin indicate two porin species at 32 and 36 kDa [51], which were summed in panel C. The values are means ± SE for six rats. Different superscripts indicate a significant difference between two groups (p

    Journal: PLoS ONE

    Article Title: Dietary Fat Influences the Expression of Contractile and Metabolic Genes in Rat Skeletal Muscle

    doi: 10.1371/journal.pone.0080152

    Figure Lengend Snippet: Gene and protein expression in the EDL muscle (A–C). (A) Relative expression levels of genes for energy metabolism (myoglobin, LPL, UCP3, PDK4), mitochondrial proteins (porin, MTCO2, UCP3 and PDK4), and transcription factors (PPARδ, FOXO1, MyoD, myogenin) and a coactivator (PGC1α) for muscle fiber type regulation in EDL muscles from soybean oil-fed (open bars), fish oil-fed (filled bars), and lard-fed (gray bars) rats. Each gene was normalized to β-actin mRNA. Values are expressed as fold changes compared with the soybean oil-fed group. (B) Protein expression levels of fiber type-related markers (myoglobin, UCP3, porin, PGC1α, and FOXO1) and loading control (actin) in the EDL, with densitometry quantification in panel C. The two arrowheads in porin indicate two porin species at 32 and 36 kDa [51], which were summed in panel C. The values are means ± SE for six rats. Different superscripts indicate a significant difference between two groups (p

    Article Snippet: The following antibodies were used: mouse monoclonal anti-myoglobin (Sigma M7773, 1:1000); anti-pan-actin (Chemicon MAB1501 (clone C4), 1:3000000); anti-porin (Abcam ab14734, 1:4000000); rabbit polyclonal anti-UCP3 (Abcam ab3477, 1:20000); anti-PGC1α (Calbiochem 516557, 1:10000); and rabbit monoclonal anti-FOXO1 (Epitomics 1874-1, 1:2000).

    Techniques: Expressing, Fluorescence In Situ Hybridization

    Gene and protein expression in the soleus muscle (A–C). (A) Relative expression levels of genes for energy metabolism (myoglobin, LPL, UCP3, PDK4), mitochondrial proteins (porin, MTCO2, UCP3 and PDK4), and transcription factors (PPARδ, FOXO1, MyoD, myogenin) and a coactivator (PGC1α) for muscle fiber type regulation in soleus muscles from soybean oil-fed (open bars), fish oil-fed (filled bars), and lard-fed (gray bars) rats. Each gene was normalized to β-actin mRNA. Values are expressed as fold changes compared with the soybean oil-fed group. (B) Protein expression levels of fiber type-related markers (myoglobin, UCP3, porin, PGC1α, and FOXO1) and loading control (actin) in the soleus, with densitometry quantification in panel C. The two arrowheads in porin indicate two porin species at 32 and 36 kDa [51], which were summed in panel C. The values are means ± SE for six rats. Different superscripts indicate a significant difference between two groups (p

    Journal: PLoS ONE

    Article Title: Dietary Fat Influences the Expression of Contractile and Metabolic Genes in Rat Skeletal Muscle

    doi: 10.1371/journal.pone.0080152

    Figure Lengend Snippet: Gene and protein expression in the soleus muscle (A–C). (A) Relative expression levels of genes for energy metabolism (myoglobin, LPL, UCP3, PDK4), mitochondrial proteins (porin, MTCO2, UCP3 and PDK4), and transcription factors (PPARδ, FOXO1, MyoD, myogenin) and a coactivator (PGC1α) for muscle fiber type regulation in soleus muscles from soybean oil-fed (open bars), fish oil-fed (filled bars), and lard-fed (gray bars) rats. Each gene was normalized to β-actin mRNA. Values are expressed as fold changes compared with the soybean oil-fed group. (B) Protein expression levels of fiber type-related markers (myoglobin, UCP3, porin, PGC1α, and FOXO1) and loading control (actin) in the soleus, with densitometry quantification in panel C. The two arrowheads in porin indicate two porin species at 32 and 36 kDa [51], which were summed in panel C. The values are means ± SE for six rats. Different superscripts indicate a significant difference between two groups (p

    Article Snippet: The following antibodies were used: mouse monoclonal anti-myoglobin (Sigma M7773, 1:1000); anti-pan-actin (Chemicon MAB1501 (clone C4), 1:3000000); anti-porin (Abcam ab14734, 1:4000000); rabbit polyclonal anti-UCP3 (Abcam ab3477, 1:20000); anti-PGC1α (Calbiochem 516557, 1:10000); and rabbit monoclonal anti-FOXO1 (Epitomics 1874-1, 1:2000).

    Techniques: Expressing, Fluorescence In Situ Hybridization

    AMPK activation on the expressions of AT 1 R and SIRT1, mitochondrial biogenesis and ROS level in RVLM of young offspring exposed to maternal HFD or ND. Representative gels and densitometric analysis of results from Western blot showing changes in protein expression of ( a ) AT 1 R; ( b ) SIRT1, ( c ) PGC-1α, or ( d ) TFAM; ( e ) mitochondrial DNA copy number; and ( f ) ROS level in RVLM of ND or HFD offspring, alone or with additional treatment with an AMPK activator, metformin (400 mg·kg − 1 ·day − 1 ), administered via gastric gavage at age of 8 weeks for 4 weeks. Analysis was performed on tissues collected bilaterally from individual RVLM at age of 12 weeks. Data on protein expression were normalized to the average ND control value, which is set to 1.0, and are presented as mean ± SEM ( n = 8 in all groups). * P

    Journal: Journal of Biomedical Science

    Article Title: Anomalous AMPK-regulated angiotensin AT1R expression and SIRT1-mediated mitochondrial biogenesis at RVLM in hypertension programming of offspring to maternal high fructose exposure

    doi: 10.1186/s12929-020-00660-z

    Figure Lengend Snippet: AMPK activation on the expressions of AT 1 R and SIRT1, mitochondrial biogenesis and ROS level in RVLM of young offspring exposed to maternal HFD or ND. Representative gels and densitometric analysis of results from Western blot showing changes in protein expression of ( a ) AT 1 R; ( b ) SIRT1, ( c ) PGC-1α, or ( d ) TFAM; ( e ) mitochondrial DNA copy number; and ( f ) ROS level in RVLM of ND or HFD offspring, alone or with additional treatment with an AMPK activator, metformin (400 mg·kg − 1 ·day − 1 ), administered via gastric gavage at age of 8 weeks for 4 weeks. Analysis was performed on tissues collected bilaterally from individual RVLM at age of 12 weeks. Data on protein expression were normalized to the average ND control value, which is set to 1.0, and are presented as mean ± SEM ( n = 8 in all groups). * P

    Article Snippet: The primary antisera used included goat polyclonal, rabbit polyclonal or monoclonal, or mouse monoclonal antiserum against AT1 R (1:2000; Santa Cruz Biotechnology, Santa Cruz, CA), gp91phox (1:5000; BD Biosciences, Sparks, MD), p67phox or p47phox (1:5000; Santa Cruz Biotechnology), manganese SOD (SOD1, 1:6000; Stressgen, Ann Arbor, MI), copper/zinc SOD (SOD2, 1:3000; Stressgen), extracellular SOD (SOD3, 1:5000; Stressgen), catalase (1:4000; Stressgen), GPx1 or GPx 3 (1:5000; BD Biosciences), NOS1, NOS2 or NOS3 (1:2000; BD Biosciences), t-AMPKα or p-AMPKα at Thr172 (1:1000; Cell Signaling, Danvers, MA), SIRT1 or SIRT3 (1:1000; Proteintech, Rosemont, IL), PGC-1α (1:1000; Santa Cruz Biotechnology), TFAM (1:1000; Abcam, Rockville, MA), leptin (1:1000; Abcam) or GAPDH (1:10,000; Merck).

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

    Schematic depiction of the contribution of AMPK/SIRT1 signaling in RVLM to oxidative stress-associated programming of hypertension in young offspring exposed to maternal high fructose. Circulatory dyslipidemia programmed by maternal HFD exposure during gestation and lactation increases leptin level, which may in turn increase AT 1 R expression and inhibit AMPK phosphorylation in RVLM. AT 1 R overexpression leads to tissue oxidative stress via an increase in gp91 phox and decrease in SOD2 expression. Suppressed AMPK activation results in inhibition of SIRT1 expression and its downstream signals, PGC-1α and TFAM, followed by tissue oxidative stress through the reduction in mitochondrial biogenesis. Suppressed AMPK activation also augments AT 1 R expression that promotes further oxidative stress. Accumulated ROS in RVLM contributes to the programmed hypertension in young HFD offspring via sympathoexcitation. The hypertension programming in HFD offspring could be protected by treatments with simvastatin to reduce dyslipidemia and hyperleptinemia; metformin to activate the AMPK/SIRT1 signals, and losartan to antagonize AT 1 R activation in RVLM. Arrows indicate activation; bar-headed lines indicate inhibition. Solid line denotes data from the present study or existing literature; dotted line denotes potential connections that require further documentation

    Journal: Journal of Biomedical Science

    Article Title: Anomalous AMPK-regulated angiotensin AT1R expression and SIRT1-mediated mitochondrial biogenesis at RVLM in hypertension programming of offspring to maternal high fructose exposure

    doi: 10.1186/s12929-020-00660-z

    Figure Lengend Snippet: Schematic depiction of the contribution of AMPK/SIRT1 signaling in RVLM to oxidative stress-associated programming of hypertension in young offspring exposed to maternal high fructose. Circulatory dyslipidemia programmed by maternal HFD exposure during gestation and lactation increases leptin level, which may in turn increase AT 1 R expression and inhibit AMPK phosphorylation in RVLM. AT 1 R overexpression leads to tissue oxidative stress via an increase in gp91 phox and decrease in SOD2 expression. Suppressed AMPK activation results in inhibition of SIRT1 expression and its downstream signals, PGC-1α and TFAM, followed by tissue oxidative stress through the reduction in mitochondrial biogenesis. Suppressed AMPK activation also augments AT 1 R expression that promotes further oxidative stress. Accumulated ROS in RVLM contributes to the programmed hypertension in young HFD offspring via sympathoexcitation. The hypertension programming in HFD offspring could be protected by treatments with simvastatin to reduce dyslipidemia and hyperleptinemia; metformin to activate the AMPK/SIRT1 signals, and losartan to antagonize AT 1 R activation in RVLM. Arrows indicate activation; bar-headed lines indicate inhibition. Solid line denotes data from the present study or existing literature; dotted line denotes potential connections that require further documentation

    Article Snippet: The primary antisera used included goat polyclonal, rabbit polyclonal or monoclonal, or mouse monoclonal antiserum against AT1 R (1:2000; Santa Cruz Biotechnology, Santa Cruz, CA), gp91phox (1:5000; BD Biosciences, Sparks, MD), p67phox or p47phox (1:5000; Santa Cruz Biotechnology), manganese SOD (SOD1, 1:6000; Stressgen, Ann Arbor, MI), copper/zinc SOD (SOD2, 1:3000; Stressgen), extracellular SOD (SOD3, 1:5000; Stressgen), catalase (1:4000; Stressgen), GPx1 or GPx 3 (1:5000; BD Biosciences), NOS1, NOS2 or NOS3 (1:2000; BD Biosciences), t-AMPKα or p-AMPKα at Thr172 (1:1000; Cell Signaling, Danvers, MA), SIRT1 or SIRT3 (1:1000; Proteintech, Rosemont, IL), PGC-1α (1:1000; Santa Cruz Biotechnology), TFAM (1:1000; Abcam, Rockville, MA), leptin (1:1000; Abcam) or GAPDH (1:10,000; Merck).

    Techniques: Expressing, Over Expression, Activation Assay, Inhibition, Pyrolysis Gas Chromatography

    Mitochondrial biogenesis in RVLM of young offspring exposed to maternal HFD or ND. Representative gels (insets) and densitometric analysis of results from Western blot showing changes in protein expression of ( a ) PGC-1α ( n = 10 per group) and ( b ) TFAM ( n = 8 per group), two key molecules in transcription and replication of mitochondrial DNA; ( c ) mitochondrial DNA copy number ( n = 10 per group); and ( d ) tissue content of ATP ( n = 6 per group) in RVLM of ND or HFD offspring, alone or with additional treatment with simvastatin (5 mg·kg − 1 ·day − 1 ), administered via gastric gavage at age of 8 weeks for 4 weeks. Analysis was performed on tissues collected bilaterally from individual RVLM at age of 12 weeks. Data on protein expression were normalized to the average ND control value, which is set to 1.0, are presented as mean ± SEM. * P

    Journal: Journal of Biomedical Science

    Article Title: Anomalous AMPK-regulated angiotensin AT1R expression and SIRT1-mediated mitochondrial biogenesis at RVLM in hypertension programming of offspring to maternal high fructose exposure

    doi: 10.1186/s12929-020-00660-z

    Figure Lengend Snippet: Mitochondrial biogenesis in RVLM of young offspring exposed to maternal HFD or ND. Representative gels (insets) and densitometric analysis of results from Western blot showing changes in protein expression of ( a ) PGC-1α ( n = 10 per group) and ( b ) TFAM ( n = 8 per group), two key molecules in transcription and replication of mitochondrial DNA; ( c ) mitochondrial DNA copy number ( n = 10 per group); and ( d ) tissue content of ATP ( n = 6 per group) in RVLM of ND or HFD offspring, alone or with additional treatment with simvastatin (5 mg·kg − 1 ·day − 1 ), administered via gastric gavage at age of 8 weeks for 4 weeks. Analysis was performed on tissues collected bilaterally from individual RVLM at age of 12 weeks. Data on protein expression were normalized to the average ND control value, which is set to 1.0, are presented as mean ± SEM. * P

    Article Snippet: The primary antisera used included goat polyclonal, rabbit polyclonal or monoclonal, or mouse monoclonal antiserum against AT1 R (1:2000; Santa Cruz Biotechnology, Santa Cruz, CA), gp91phox (1:5000; BD Biosciences, Sparks, MD), p67phox or p47phox (1:5000; Santa Cruz Biotechnology), manganese SOD (SOD1, 1:6000; Stressgen, Ann Arbor, MI), copper/zinc SOD (SOD2, 1:3000; Stressgen), extracellular SOD (SOD3, 1:5000; Stressgen), catalase (1:4000; Stressgen), GPx1 or GPx 3 (1:5000; BD Biosciences), NOS1, NOS2 or NOS3 (1:2000; BD Biosciences), t-AMPKα or p-AMPKα at Thr172 (1:1000; Cell Signaling, Danvers, MA), SIRT1 or SIRT3 (1:1000; Proteintech, Rosemont, IL), PGC-1α (1:1000; Santa Cruz Biotechnology), TFAM (1:1000; Abcam, Rockville, MA), leptin (1:1000; Abcam) or GAPDH (1:10,000; Merck).

    Techniques: Western Blot, Expressing, Pyrolysis Gas Chromatography

    Involvement of the PGC1α/Nrf2 pathway in neuroprotection exerted by compound 22a in CGNs. (A) Representative blots showed the expression of proteins PGC1α, Nrf2 and HO-1 in CGNs after different treatments. (B) Densitometric analysis of the protein expression in (A) . (C) Representative blots showed the protein expression of PGC1α was silenced in CGNs pretreated with PGC1α siRNA, but not when pretreated with scrambled siRNA. (D) Densitometric analysis of (C) . (E) PGC1α siRNA significantly attenuated the neuroprotective effects of compound 22a against glutamate-induced neuronal cell death in CGNs. CGNs were transfected with either PGC1α siRNA or scrambled siRNA for 24 h and then pretreated with compound 22a (10 mM) for 2 h before exposure to 200 μM glutamate. Cell viability was measured at 24 h post glutamate exposure using the MTT assay. Data were expressed as the mean ± SEM of three experiments; ### p

    Journal: Frontiers in Neuroscience

    Article Title: A Novel Tetramethylpyrazine Derivative Protects Against Glutamate-Induced Cytotoxicity Through PGC1α/Nrf2 and PI3K/Akt Signaling Pathways

    doi: 10.3389/fnins.2018.00567

    Figure Lengend Snippet: Involvement of the PGC1α/Nrf2 pathway in neuroprotection exerted by compound 22a in CGNs. (A) Representative blots showed the expression of proteins PGC1α, Nrf2 and HO-1 in CGNs after different treatments. (B) Densitometric analysis of the protein expression in (A) . (C) Representative blots showed the protein expression of PGC1α was silenced in CGNs pretreated with PGC1α siRNA, but not when pretreated with scrambled siRNA. (D) Densitometric analysis of (C) . (E) PGC1α siRNA significantly attenuated the neuroprotective effects of compound 22a against glutamate-induced neuronal cell death in CGNs. CGNs were transfected with either PGC1α siRNA or scrambled siRNA for 24 h and then pretreated with compound 22a (10 mM) for 2 h before exposure to 200 μM glutamate. Cell viability was measured at 24 h post glutamate exposure using the MTT assay. Data were expressed as the mean ± SEM of three experiments; ### p

    Article Snippet: Antibodies against β-actin, PGC1α, Nrf2 and HO-1 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, United States).

    Techniques: Expressing, Transfection, MTT Assay

    Compound 22a activates of PGC1α/Nrf2 signaling pathway. (A) Compound 22a (0.1, 1, 10, 100 μM) increased the transcriptional activity of PGC1α determined by the luciferase reporter assay. (B) Compound 22a (100 μM) induced the nuclear translocation of Nrf2 determined by immunofluorescence staining. Left panel: green fluorescence showing Nrf2 localization. Middle panel: stained nucleus with DAPI. Right panel: merged images from green and blue filters. Scale bar: 20 μm. (C) Immunoblot assay was performed with antibodies against Nrf2 and HO-1. (D,E) Densitometric analysis of the protein expression in (C) . CGNs were treated with compound 22a (1, 10, 100 μM) for 12 h. Data were expressed as the mean ± SEM of three separate experiments; ∗ p

    Journal: Frontiers in Neuroscience

    Article Title: A Novel Tetramethylpyrazine Derivative Protects Against Glutamate-Induced Cytotoxicity Through PGC1α/Nrf2 and PI3K/Akt Signaling Pathways

    doi: 10.3389/fnins.2018.00567

    Figure Lengend Snippet: Compound 22a activates of PGC1α/Nrf2 signaling pathway. (A) Compound 22a (0.1, 1, 10, 100 μM) increased the transcriptional activity of PGC1α determined by the luciferase reporter assay. (B) Compound 22a (100 μM) induced the nuclear translocation of Nrf2 determined by immunofluorescence staining. Left panel: green fluorescence showing Nrf2 localization. Middle panel: stained nucleus with DAPI. Right panel: merged images from green and blue filters. Scale bar: 20 μm. (C) Immunoblot assay was performed with antibodies against Nrf2 and HO-1. (D,E) Densitometric analysis of the protein expression in (C) . CGNs were treated with compound 22a (1, 10, 100 μM) for 12 h. Data were expressed as the mean ± SEM of three separate experiments; ∗ p

    Article Snippet: Antibodies against β-actin, PGC1α, Nrf2 and HO-1 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, United States).

    Techniques: Activity Assay, Luciferase, Reporter Assay, Translocation Assay, Immunofluorescence, Staining, Fluorescence, Expressing

    Western blot analyses of protein expression and RT-PCR analysis of mitochondria-related gene expression in ALA + Fe treated cisplatin-induced renal tubular injury. (A) Aliquots of 50 μg of protein extracts from NRK-52E cells were separated by SDS-PAGE and transferred to membranes. Western blots analyses were performed for ATP5α, complex (COX)-IV, PGC-1α, UCP2 in control, cisplatin, cisplatin + ALA, and cisplatin + ALA + Fe treated NRK-52E cells. Actin served as a loading control. (B) Quantitative densitometry was performed for ATP5α, complex (COX)-IV, PGC-1α, and UCP2 blots. (C) Quantitative analysis of mRNA was performed using RT-PCR for ATP5α, complex (COX)-IV, PGC-1α, and UCP2. Bars represent the mean ± SEM, n = 6. *p

    Journal: PLoS ONE

    Article Title: 5-Aminolevulinic Acid Protects against Cisplatin-Induced Nephrotoxicity without Compromising the Anticancer Efficiency of Cisplatin in Rats In Vitro and In Vivo

    doi: 10.1371/journal.pone.0080850

    Figure Lengend Snippet: Western blot analyses of protein expression and RT-PCR analysis of mitochondria-related gene expression in ALA + Fe treated cisplatin-induced renal tubular injury. (A) Aliquots of 50 μg of protein extracts from NRK-52E cells were separated by SDS-PAGE and transferred to membranes. Western blots analyses were performed for ATP5α, complex (COX)-IV, PGC-1α, UCP2 in control, cisplatin, cisplatin + ALA, and cisplatin + ALA + Fe treated NRK-52E cells. Actin served as a loading control. (B) Quantitative densitometry was performed for ATP5α, complex (COX)-IV, PGC-1α, and UCP2 blots. (C) Quantitative analysis of mRNA was performed using RT-PCR for ATP5α, complex (COX)-IV, PGC-1α, and UCP2. Bars represent the mean ± SEM, n = 6. *p

    Article Snippet: The membranes were blocked for 1 h with 5% (wt/vol) fat-free milk in PBS and probed with the appropriate primary antibodies (anti-ATP5α, anti-complex [COX]-IV, anti-PGC-1α, anti-UCP2, anti-nitrotyrosine [anti-NT], anti-procaspase-3, or anti-actin [Santa Cruz Biochemicals Inc., Santa Cruz, CA, USA]).

    Techniques: Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, SDS Page, Pyrolysis Gas Chromatography

    Western blot analyses of protein expression and RT-PCR analysis of mitochondria-related gene expression in ALA treated cisplatin-induced AKI rats. (A) Aliquots of 50 μg of protein from renal tissue extracts were separated by SDS-PAGE and transferred to membranes. Western blots analyses were performed for ATP5α, complex (COX)-IV, PGC-1α, UCP2 in cisplatin-treated, control, and cisplatin + ALA (both post and pre post) treated rats. Actin served as a loading control. (B) Quantitative densitometry was performed for ATP5α, complex (COX)-IV, PGC-1α, and UCP2 western blots. (C) Quantitative analysis of mRNA was performed using RT-PCR for ATP5α, complex (COX)-IV, PGC-1α, and UCP2. GAPDH served as a loading control. Bars represent the mean ± SEM, n = 6. *p

    Journal: PLoS ONE

    Article Title: 5-Aminolevulinic Acid Protects against Cisplatin-Induced Nephrotoxicity without Compromising the Anticancer Efficiency of Cisplatin in Rats In Vitro and In Vivo

    doi: 10.1371/journal.pone.0080850

    Figure Lengend Snippet: Western blot analyses of protein expression and RT-PCR analysis of mitochondria-related gene expression in ALA treated cisplatin-induced AKI rats. (A) Aliquots of 50 μg of protein from renal tissue extracts were separated by SDS-PAGE and transferred to membranes. Western blots analyses were performed for ATP5α, complex (COX)-IV, PGC-1α, UCP2 in cisplatin-treated, control, and cisplatin + ALA (both post and pre post) treated rats. Actin served as a loading control. (B) Quantitative densitometry was performed for ATP5α, complex (COX)-IV, PGC-1α, and UCP2 western blots. (C) Quantitative analysis of mRNA was performed using RT-PCR for ATP5α, complex (COX)-IV, PGC-1α, and UCP2. GAPDH served as a loading control. Bars represent the mean ± SEM, n = 6. *p

    Article Snippet: The membranes were blocked for 1 h with 5% (wt/vol) fat-free milk in PBS and probed with the appropriate primary antibodies (anti-ATP5α, anti-complex [COX]-IV, anti-PGC-1α, anti-UCP2, anti-nitrotyrosine [anti-NT], anti-procaspase-3, or anti-actin [Santa Cruz Biochemicals Inc., Santa Cruz, CA, USA]).

    Techniques: Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, SDS Page, Pyrolysis Gas Chromatography

    Aberrant mitochondrial fission and altered expression of fission regulatory proteins in D7‐Des Tg hearts. A, Representative transmission electron micrographs of hearts from D7‐Des Tg mice (Scale bars: 2 μm). B, Relative mtDNA content expressed compared with total genomic DNA (nDNA) in hearts of D7‐Des Tg was significantly increased compared with Ntg. Representative Western blot and densitometric quantification of the (C) whole cell fraction and (D) mitochondrial fraction showing expression of mitochondrial dynamic regulatory proteins in the D7‐Des Tg hearts: Drp1, pDrp1 (Ser 616), Fis 1, OPA1, OMA1, MFN2, Mff, Tom20, Tim23, and PGC1α. Ponceau S protein staining of the transfer membrane confirmed approximately equal loading across the gel. Bars represent mean±SEM. n = 6 mice per group. P values were determined by Tukey's post hoc test. D7‐Des Tg indicates mutant desmin transgenic mouse; mtDNA, mitochondrial DNA; NS, not significant; Ntg, non‐transgenic.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Aberrant Mitochondrial Fission Is Maladaptive in Desmin Mutation–Induced Cardiac Proteotoxicity

    doi: 10.1161/JAHA.118.009289

    Figure Lengend Snippet: Aberrant mitochondrial fission and altered expression of fission regulatory proteins in D7‐Des Tg hearts. A, Representative transmission electron micrographs of hearts from D7‐Des Tg mice (Scale bars: 2 μm). B, Relative mtDNA content expressed compared with total genomic DNA (nDNA) in hearts of D7‐Des Tg was significantly increased compared with Ntg. Representative Western blot and densitometric quantification of the (C) whole cell fraction and (D) mitochondrial fraction showing expression of mitochondrial dynamic regulatory proteins in the D7‐Des Tg hearts: Drp1, pDrp1 (Ser 616), Fis 1, OPA1, OMA1, MFN2, Mff, Tom20, Tim23, and PGC1α. Ponceau S protein staining of the transfer membrane confirmed approximately equal loading across the gel. Bars represent mean±SEM. n = 6 mice per group. P values were determined by Tukey's post hoc test. D7‐Des Tg indicates mutant desmin transgenic mouse; mtDNA, mitochondrial DNA; NS, not significant; Ntg, non‐transgenic.

    Article Snippet: Santa Cruz Biotechnology), Anti‐Fis1 (1:200, Santa Cruz Biotechnology), Anti‐mitochondrial fission factor (1:200, Santa Cruz Biotechnology), Anti‐Tom20 (1:200, Santa Cruz Biotechnology), Anti‐Tim23 (1:200, Santa Cruz Biotechnology), Anti‐PGC1α (1:200, Santa Cruz Biotechnology), Anti‐β‐Actin (1:1000, Santa Cruz Biotechnology), Anti‐OXPHOS (1:1000, Abcam), Anti–pyruvate dehydrogenase (1:1000, Abcam), Anti‐Bax (1:1000, Cell Signaling Technology), Anti‐COXIV (1:1000, Cell Signaling Technology), Anti‐cleaved PARP1 (1:200, Santa Cruz Biotechnology), Anti‐CHOP (1:1000, Cell Signaling Technology), Anti‐P62 (Progen Biotechnik GmbH), Anti‐Desmin (1:2000, Biomeda), Anti‐CryAB (1:10 000, Enzo Life Sciences), Anti‐GRP75 (1:200, Santa Cruz Biotechnology), Anti‐Periostin (1:500, Santa Cruz Biotechnology), and β‐Tubulin (1:500) (Cell Signaling Technology).

    Techniques: Expressing, Transmission Assay, Mouse Assay, Western Blot, Staining, Mutagenesis, Transgenic Assay

    Mitochondrial fission inhibition by mdivi‐1 in D7‐Des overexpressed NRCs. A, Representative Western blot and densitometric quantification showing Drp1, OPA1, and PGC1α expression after 5 μmol/L mdivi‐1 or vehicle for 24 hours in D7‐Des overexpressed cardiomyocytes. B, Expression of Bax and CHOP in the whole cell lysate, and (C) LDH release in the medium after 5 μmol/L mdivi‐1 or vehicle treatment. D, Desmin expression in the pellet fraction was significantly decreased after mdivi‐1 treatment. E, Desmin expression in the whole cell fraction was not significantly changed after mdivi‐1 treatment. Ponceau S protein stain of the transfer membrane was used to confirm approximately equal loading. n=3 replicates per group. Boxes represents interquartile ranges, lines represent medians, whiskers represent ranges, and P values were determined by Kruskal–Wallis test. LDH indicates lactate dehydrogenase; MOI, multiplicities of infection; NRCs, neonatal rat cardiomyocytes; NS, not significant.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Aberrant Mitochondrial Fission Is Maladaptive in Desmin Mutation–Induced Cardiac Proteotoxicity

    doi: 10.1161/JAHA.118.009289

    Figure Lengend Snippet: Mitochondrial fission inhibition by mdivi‐1 in D7‐Des overexpressed NRCs. A, Representative Western blot and densitometric quantification showing Drp1, OPA1, and PGC1α expression after 5 μmol/L mdivi‐1 or vehicle for 24 hours in D7‐Des overexpressed cardiomyocytes. B, Expression of Bax and CHOP in the whole cell lysate, and (C) LDH release in the medium after 5 μmol/L mdivi‐1 or vehicle treatment. D, Desmin expression in the pellet fraction was significantly decreased after mdivi‐1 treatment. E, Desmin expression in the whole cell fraction was not significantly changed after mdivi‐1 treatment. Ponceau S protein stain of the transfer membrane was used to confirm approximately equal loading. n=3 replicates per group. Boxes represents interquartile ranges, lines represent medians, whiskers represent ranges, and P values were determined by Kruskal–Wallis test. LDH indicates lactate dehydrogenase; MOI, multiplicities of infection; NRCs, neonatal rat cardiomyocytes; NS, not significant.

    Article Snippet: Santa Cruz Biotechnology), Anti‐Fis1 (1:200, Santa Cruz Biotechnology), Anti‐mitochondrial fission factor (1:200, Santa Cruz Biotechnology), Anti‐Tom20 (1:200, Santa Cruz Biotechnology), Anti‐Tim23 (1:200, Santa Cruz Biotechnology), Anti‐PGC1α (1:200, Santa Cruz Biotechnology), Anti‐β‐Actin (1:1000, Santa Cruz Biotechnology), Anti‐OXPHOS (1:1000, Abcam), Anti–pyruvate dehydrogenase (1:1000, Abcam), Anti‐Bax (1:1000, Cell Signaling Technology), Anti‐COXIV (1:1000, Cell Signaling Technology), Anti‐cleaved PARP1 (1:200, Santa Cruz Biotechnology), Anti‐CHOP (1:1000, Cell Signaling Technology), Anti‐P62 (Progen Biotechnik GmbH), Anti‐Desmin (1:2000, Biomeda), Anti‐CryAB (1:10 000, Enzo Life Sciences), Anti‐GRP75 (1:200, Santa Cruz Biotechnology), Anti‐Periostin (1:500, Santa Cruz Biotechnology), and β‐Tubulin (1:500) (Cell Signaling Technology).

    Techniques: Inhibition, Western Blot, Expressing, Staining, Infection

    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: After the final wash, the samples were dissolved in 100 μl of elution solution (0.1 M glycine, pH 2.5); 20 μl were then separated by SDS-PAGE and immunoblotted, as described [ ], using an acetyl-lysine antibody (1:1.000; Cell Signaling) and anti-PGC-1α antibody (1:1.000; Santa Cruz), respectively.

    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: After the final wash, the samples were dissolved in 100 μl of elution solution (0.1 M glycine, pH 2.5); 20 μl were then separated by SDS-PAGE and immunoblotted, as described [ ], using an acetyl-lysine antibody (1:1.000; Cell Signaling) and anti-PGC-1α antibody (1:1.000; Santa Cruz), respectively.

    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: After the final wash, the samples were dissolved in 100 μl of elution solution (0.1 M glycine, pH 2.5); 20 μl were then separated by SDS-PAGE and immunoblotted, as described [ ], using an acetyl-lysine antibody (1:1.000; Cell Signaling) and anti-PGC-1α antibody (1:1.000; Santa Cruz), respectively.

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

    RN-1 induces γ-globin synthesis by inducing PGC-1α. (A-B) QRT-PCR analyses quantify the fold change in LSD1 and PGC-1α mRNAs after normalization to the expression of Oaz1. (C) Diagrammatic representation of the gain-of-function

    Journal: Blood

    Article Title: The LSD1 inhibitor RN-1 induces fetal hemoglobin synthesis and reduces disease pathology in sickle cell mice

    doi: 10.1182/blood-2015-02-626259

    Figure Lengend Snippet: RN-1 induces γ-globin synthesis by inducing PGC-1α. (A-B) QRT-PCR analyses quantify the fold change in LSD1 and PGC-1α mRNAs after normalization to the expression of Oaz1. (C) Diagrammatic representation of the gain-of-function

    Article Snippet: Protein extracts from control or RN-1–treated murine erythroleukemia (MEL) cells or SCD mice BM cells were subjected to western blotting as described, using the same antibodies against LSD1, H3K4me2, H3K9me2, and H3, as well as antibodies against PGC-1α and β-actin (Santa Cruz Biotechnology).

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

    PGC-1α, PPARα and GyK expression increases in KO mice. EATs were dissected from miR-150 KO (n = 7) and WT (n = 7) mice after 17 weeks HFD feeding. A) PGC-1α, PPARα and GyK gene expression was assessed by qRT-PCR. B) Protein levels of PGC-1α and GyK were measured by Western blotting. C) Western blotting results were quantified by ImageJ. PGC-1α and GyK expression were normalized by β-actin and GAPDH respectively. Data represent mean ± SEM, *p

    Journal: Metabolism: clinical and experimental

    Article Title: Improved systemic metabolism and adipocyte biology in miR-150 knockout mice

    doi: 10.1016/j.metabol.2017.12.018

    Figure Lengend Snippet: PGC-1α, PPARα and GyK expression increases in KO mice. EATs were dissected from miR-150 KO (n = 7) and WT (n = 7) mice after 17 weeks HFD feeding. A) PGC-1α, PPARα and GyK gene expression was assessed by qRT-PCR. B) Protein levels of PGC-1α and GyK were measured by Western blotting. C) Western blotting results were quantified by ImageJ. PGC-1α and GyK expression were normalized by β-actin and GAPDH respectively. Data represent mean ± SEM, *p

    Article Snippet: The membrane was blocked with 5% milk for 1 h at room temperature (RT) and exposed to the primary anti-bodies overnight at 4 °C (mTOR, Cell signaling cat#2983; pAKT, Cell signaling cat#9271; pIRS-1, Santa Cruz cat#sc17196; PGC-1α, Abcam cat#ab54481; and GyK, Abcam cat#ab126599).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Mouse Assay, Quantitative RT-PCR, Western Blot

    Rescuing mGPDH deficiency improves skeletal muscle regeneration during obesity and diabetes A–C Immunoblot (A, C) and IHC (B) of mGPDH and myogenin in GA muscles from obese patients (A, B) and the indicated mice (C). D qRT–PCR of mGPDH in GA muscle of the indicated mice at days 0 and 3 after CTX intramuscular injection. E–H Experimental setup (E, upper panel); qRT–PCR of mGPDH, myogenin, and myh3 (E, bottom panel); H E staining (arrowhead, necrotic myofibers; asterisks, regenerating fibers) (F); distribution of the fiber CSA (G); and percentage of myofibers with central nuclei (H) in GA muscle from AAV‐mGPDH‐treated HFD‐fed mice at day 7 after CTX intramuscular injection. I–M Experimental setup (I and M, upper panels); qRT–PCR of mGPDH, myogenin, and myh3 (I, bottom panel); H E staining (arrowhead, necrotic myofibers; asterisks, regenerating fibers) (J); distribution of the fibers CSA (K); percentage of myofibers with central nuclei (L); and muscle weight (M, bottom panel) in GA muscle from AAV‐mGPDH‐treated STZ‐treated mice at days 7 (I–L) and 14 (M) after CTX intramuscular injection. N Immunoblots of mGPDH, p‐AMPK, p‐ACC, PGC1α, and myogenin for the experiment described in (E). Data information: Data are presented as the mean ± s.e.m. Scale bars represent 200 μm in panel (B) and 100 μm (25 μm for magnification insets) in panels (F, J). In panels (A, B), obese patients ( n = 11) and normal subjects ( n = 18); in panels (C, D), n = 3 mice per group; in panels (E–L and N), n = 6 mice per group; in panel (M), n = 4 mice per group; in panels (B, F–H, and J–L), three sections were obtained per mouse. * P

    Journal: EMBO Molecular Medicine

    Article Title: Mitochondrial glycerol 3‐phosphate dehydrogenase promotes skeletal muscle regeneration

    doi: 10.15252/emmm.201809390

    Figure Lengend Snippet: Rescuing mGPDH deficiency improves skeletal muscle regeneration during obesity and diabetes A–C Immunoblot (A, C) and IHC (B) of mGPDH and myogenin in GA muscles from obese patients (A, B) and the indicated mice (C). D qRT–PCR of mGPDH in GA muscle of the indicated mice at days 0 and 3 after CTX intramuscular injection. E–H Experimental setup (E, upper panel); qRT–PCR of mGPDH, myogenin, and myh3 (E, bottom panel); H E staining (arrowhead, necrotic myofibers; asterisks, regenerating fibers) (F); distribution of the fiber CSA (G); and percentage of myofibers with central nuclei (H) in GA muscle from AAV‐mGPDH‐treated HFD‐fed mice at day 7 after CTX intramuscular injection. I–M Experimental setup (I and M, upper panels); qRT–PCR of mGPDH, myogenin, and myh3 (I, bottom panel); H E staining (arrowhead, necrotic myofibers; asterisks, regenerating fibers) (J); distribution of the fibers CSA (K); percentage of myofibers with central nuclei (L); and muscle weight (M, bottom panel) in GA muscle from AAV‐mGPDH‐treated STZ‐treated mice at days 7 (I–L) and 14 (M) after CTX intramuscular injection. N Immunoblots of mGPDH, p‐AMPK, p‐ACC, PGC1α, and myogenin for the experiment described in (E). Data information: Data are presented as the mean ± s.e.m. Scale bars represent 200 μm in panel (B) and 100 μm (25 μm for magnification insets) in panels (F, J). In panels (A, B), obese patients ( n = 11) and normal subjects ( n = 18); in panels (C, D), n = 3 mice per group; in panels (E–L and N), n = 6 mice per group; in panel (M), n = 4 mice per group; in panels (B, F–H, and J–L), three sections were obtained per mouse. * P

    Article Snippet: The following antibodies were used: mGPDH (1:1,000, sc‐390830), Myogenin (1:1,000, sc‐12732), cGPDH (1:500, sc‐376219), IGF‐1R (1:1,000, sc‐81464), GAPDH (1:10,000, sc‐20357), β‐actin (1:2,000, sc‐47778), and c‐myc (1:1,000, sc‐42) from Santa Cruz Biotechnology; MyHC (1:2,000, M4276) from Sigma; Akt (1:1,000, #4691), p‐Akt (Thr308, 1:1,000, #13038), IRS1 (1:1,000, #2382), p‐IRS1 (Ser307, 1:1,000, #2381), VDAC (1:1,000, #4866), Cyt c (1:1,000, #4280), AMPKα (1:2,000, #2532), p‐AMPKα (Thr172, 1:2,000, #2535), ACC (1:1,000, #3676), p‐ACC (Ser79, 1:1,000, #11818), Flag (1:1,000, #8146), and LC3B (1:1,000, #2775) from Cell Signaling Technology; PGC1α (1:1,000, ab54481) and mGPDH (1:10,000, ab188585) from Abcam; and COX IV (1:500, AC610) from Beyotime.

    Techniques: Immunohistochemistry, Mouse Assay, Quantitative RT-PCR, Injection, Staining, Western Blot

    mGPDH effect occurs via the CaMKKβ/AMPK control of mitochondrial biogenesis A–F Mitochondrial DNA (A), nuclear‐encoded OXPHOS genes (B), respirometry analysis (C), and immunoblots of mGPDH, phospho‐Thr172 AMPK (p‐AMPK), total AMPK (AMPK), phospho‐Ser79‐ACC (p‐ACC), total ACC and PGC1α, and corresponding quantifications represent mGPDH, p‐AMPK, p‐ACC, and PGC1α protein levels (D–F) in C2C12 myocytes transfected with siRNA or plasmid for mGPDH 24 h after differentiation. G–I Immunoblots of p‐AMPK, p‐ACC, and PGC1α and corresponding quantifications represent p‐AMPK, p‐ACC, and PGC1α protein levels (G), mitochondrial DNA (H), and nuclear‐encoded OXPHOS genes combined by NDUFS8 , SDHb , Uqcrc1 , COX5b , and ATP5a1 (I) in C2C12 myocytes transfected by mGPDH plasmid with the AMPK inhibitor compound C (CC) 24 h after differentiation. J, K NAD + /NADH ratio (J) and immunoprecipitation analysis for PGC1α acetyl‐lysine (Ac‐Lys) level (K) in C2C12 myocytes transfected with siRNA or plasmid for mGPDH 24 h after differentiation. L–P Immunoblot of c‐myc and myogenin (L) and corresponding quantifications represent c‐myc and myogenin protein levels (M), representative images of MyHC immunofluorescence (N), fusion index (O), and the distribution of nuclei per myotube (P) in C2C12 myocytes transfected with mGPDH plasmid with the AMPK inhibitor CC at 24 h (L, M) or 72 h (N–P) after differentiation. Q Immunoblots of p‐AMPK, p‐ACC, PGC1α, and myogenin in C2C12 myocytes transfected with mGPDH plasmid with the CaMKKβ inhibitor STO‐609 at 24 h after differentiation. Quantifications represent p‐AMPK, p‐ACC, PGC1α, and myogenin protein levels. R Immunoblots of p‐AMPK and p‐ACC in C2C12 myocytes transfected with mGPDH plasmid with the Ca 2+ chelator BAPTA‐AM at 24 h after differentiation. Quantifications represent p‐AMPK and p‐ACC protein levels. Data information: Data are presented as the mean ± s.e.m. Scale bars represent 50 μm in panel (N). In panels (A, B, D–M, Q, and R), n = 3; in panel (C), n = 10; in panels (N–P), n = 15. * P

    Journal: EMBO Molecular Medicine

    Article Title: Mitochondrial glycerol 3‐phosphate dehydrogenase promotes skeletal muscle regeneration

    doi: 10.15252/emmm.201809390

    Figure Lengend Snippet: mGPDH effect occurs via the CaMKKβ/AMPK control of mitochondrial biogenesis A–F Mitochondrial DNA (A), nuclear‐encoded OXPHOS genes (B), respirometry analysis (C), and immunoblots of mGPDH, phospho‐Thr172 AMPK (p‐AMPK), total AMPK (AMPK), phospho‐Ser79‐ACC (p‐ACC), total ACC and PGC1α, and corresponding quantifications represent mGPDH, p‐AMPK, p‐ACC, and PGC1α protein levels (D–F) in C2C12 myocytes transfected with siRNA or plasmid for mGPDH 24 h after differentiation. G–I Immunoblots of p‐AMPK, p‐ACC, and PGC1α and corresponding quantifications represent p‐AMPK, p‐ACC, and PGC1α protein levels (G), mitochondrial DNA (H), and nuclear‐encoded OXPHOS genes combined by NDUFS8 , SDHb , Uqcrc1 , COX5b , and ATP5a1 (I) in C2C12 myocytes transfected by mGPDH plasmid with the AMPK inhibitor compound C (CC) 24 h after differentiation. J, K NAD + /NADH ratio (J) and immunoprecipitation analysis for PGC1α acetyl‐lysine (Ac‐Lys) level (K) in C2C12 myocytes transfected with siRNA or plasmid for mGPDH 24 h after differentiation. L–P Immunoblot of c‐myc and myogenin (L) and corresponding quantifications represent c‐myc and myogenin protein levels (M), representative images of MyHC immunofluorescence (N), fusion index (O), and the distribution of nuclei per myotube (P) in C2C12 myocytes transfected with mGPDH plasmid with the AMPK inhibitor CC at 24 h (L, M) or 72 h (N–P) after differentiation. Q Immunoblots of p‐AMPK, p‐ACC, PGC1α, and myogenin in C2C12 myocytes transfected with mGPDH plasmid with the CaMKKβ inhibitor STO‐609 at 24 h after differentiation. Quantifications represent p‐AMPK, p‐ACC, PGC1α, and myogenin protein levels. R Immunoblots of p‐AMPK and p‐ACC in C2C12 myocytes transfected with mGPDH plasmid with the Ca 2+ chelator BAPTA‐AM at 24 h after differentiation. Quantifications represent p‐AMPK and p‐ACC protein levels. Data information: Data are presented as the mean ± s.e.m. Scale bars represent 50 μm in panel (N). In panels (A, B, D–M, Q, and R), n = 3; in panel (C), n = 10; in panels (N–P), n = 15. * P

    Article Snippet: The following antibodies were used: mGPDH (1:1,000, sc‐390830), Myogenin (1:1,000, sc‐12732), cGPDH (1:500, sc‐376219), IGF‐1R (1:1,000, sc‐81464), GAPDH (1:10,000, sc‐20357), β‐actin (1:2,000, sc‐47778), and c‐myc (1:1,000, sc‐42) from Santa Cruz Biotechnology; MyHC (1:2,000, M4276) from Sigma; Akt (1:1,000, #4691), p‐Akt (Thr308, 1:1,000, #13038), IRS1 (1:1,000, #2382), p‐IRS1 (Ser307, 1:1,000, #2381), VDAC (1:1,000, #4866), Cyt c (1:1,000, #4280), AMPKα (1:2,000, #2532), p‐AMPKα (Thr172, 1:2,000, #2535), ACC (1:1,000, #3676), p‐ACC (Ser79, 1:1,000, #11818), Flag (1:1,000, #8146), and LC3B (1:1,000, #2775) from Cell Signaling Technology; PGC1α (1:1,000, ab54481) and mGPDH (1:10,000, ab188585) from Abcam; and COX IV (1:500, AC610) from Beyotime.

    Techniques: Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation, Immunofluorescence

    The role of adenosine monophosphate-activated protein kinase (AMPK) signaling pathway in the protective effect of TLB on H 2 O 2 -induced PC12 cells injury. (A) Representative Western blot were shown for AMPK, peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and ERRα proteins. (B) Quantitation of phosphorylation of AMPK protein. The relative optical density was normalized to AMPK. (C) Quantitation of PGC-1α protein. (D) Quantitation of ERRα protein. AMPK-specific inhibitor significantly abolished the protective effect of TLB. (E) Cell viability was determined using MTT assay. (F) LDH release was determined using an LDH release assay. Data were presented as mean ± SD of three independent experiments. ** P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Trilobatin Protects Against Oxidative Injury in Neuronal PC12 Cells Through Regulating Mitochondrial ROS Homeostasis Mediated by AMPK/Nrf2/Sirt3 Signaling Pathway

    doi: 10.3389/fnmol.2018.00267

    Figure Lengend Snippet: The role of adenosine monophosphate-activated protein kinase (AMPK) signaling pathway in the protective effect of TLB on H 2 O 2 -induced PC12 cells injury. (A) Representative Western blot were shown for AMPK, peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and ERRα proteins. (B) Quantitation of phosphorylation of AMPK protein. The relative optical density was normalized to AMPK. (C) Quantitation of PGC-1α protein. (D) Quantitation of ERRα protein. AMPK-specific inhibitor significantly abolished the protective effect of TLB. (E) Cell viability was determined using MTT assay. (F) LDH release was determined using an LDH release assay. Data were presented as mean ± SD of three independent experiments. ** P

    Article Snippet: MitoSOX Red (#M36008) and Mito Tracker green probe (#M7514) were obtained from Invitrogen (Eugene, OR, USA), Compound C (#ab120843), Mitochondria isolation kit for cultured cells (#ab110170), NAD/NADH quantification kit (#ab65348), Complex I enzyme activity microplate assay kit (Colorimetric; #ab109721), ATP synthase enzyme activity microplate assay (#ab109714), BCA protein quantification kit (#ab102536), anti-GPX -1 (#ab22604), anti-Sirt 3 (#ab189860), anti-IDH2 (#ab129180), anti-forkhead boxO3a (FoxO3a, #ab23683), anti-SOD2 (acetyl K68; #ab13533), anti-SOD2 (#ab13533), anti-uncoupling protein 2 (UCP2, #ab203244), anti-AMPK (#ab80039), anti-p-AMPK (#ab133448), anti-PGC-1α (#ab54481), anti-estrogen-related receptor-α (#ab76228), anti-Nrf2 antibody (#ab137550), anti-kelchlike ECH-associated protein 1 (Keap-1, #ab139729), anti-HO-1 (#ab68477), anti-NQO1 (#ab34173) antibodies were purchased from Abcam (Cambridge, UK).

    Techniques: Western Blot, Pyrolysis Gas Chromatography, Quantitation Assay, MTT Assay, Lactate Dehydrogenase Assay

    Increased catecholamines level and iBAT thermogenesis in FoxO1 KO DAT mice. ( a ) Serum norepinephrine levels of WT and KO male and female mice. ( b ) Dopamine contents measured from the midbrain and SN samples of WT and KO male mice. ( c ) Gene expression in iBAT of WT and KO male mice. ( d ) Immunoblots for p-Creb, p-p38 MAPK, UCP1, PGC1α in iBAT. ( e ) Relative protein levels for p-Creb, p-p38 MAPK, UCP1, PGC1α in iBAT from ( d ). Normalized to total p38 MAPK or GAPDH. ( f ) Representative figures of H E staining and UCP1 staining from iBAT samples of WT and KO mice ( n =3). Scale bar, 50 μm. ( g ) Quantification of mitochondrial DNA contents from iBAT samples of WT and KO mice. ♂, male. ♀, female. Data are expressed as mean±s.e.m. (* P

    Journal: Nature Communications

    Article Title: FoxO1 in dopaminergic neurons regulates energy homeostasis and targets tyrosine hydroxylase

    doi: 10.1038/ncomms12733

    Figure Lengend Snippet: Increased catecholamines level and iBAT thermogenesis in FoxO1 KO DAT mice. ( a ) Serum norepinephrine levels of WT and KO male and female mice. ( b ) Dopamine contents measured from the midbrain and SN samples of WT and KO male mice. ( c ) Gene expression in iBAT of WT and KO male mice. ( d ) Immunoblots for p-Creb, p-p38 MAPK, UCP1, PGC1α in iBAT. ( e ) Relative protein levels for p-Creb, p-p38 MAPK, UCP1, PGC1α in iBAT from ( d ). Normalized to total p38 MAPK or GAPDH. ( f ) Representative figures of H E staining and UCP1 staining from iBAT samples of WT and KO mice ( n =3). Scale bar, 50 μm. ( g ) Quantification of mitochondrial DNA contents from iBAT samples of WT and KO mice. ♂, male. ♀, female. Data are expressed as mean±s.e.m. (* P

    Article Snippet: 11667149001, 1:1,000), GAPDH (Santa Cruz, Cat.No. sc-25778, 1:10,000), anti-Prolactin (Santa Cruz, Cat.No. sc-7805, 1:500), anti-PGC1α (Abcam, Cat.No. ab54481, 1:2,000), anti-Ucp1 (Abcam, Cat.No. ab10983, 1:10,000), anti-COMT (Abcam, Cat.No. ab126618, 1:1,000), anti-MAO-b (Abcam, Cat.No. ab125010, 1:2,000) and β-Actin (Abcam, Cat.No. ab6276, 1:10,000).

    Techniques: Mouse Assay, Expressing, Western Blot, Staining

    Effects of PGC-1α Overexpression in Surf1 −/− Mice (A) Expression analysis of PGC-1α . (Left panel) The PGC-1α transcript from muscle of each mouse genotype (three animals/genotype) was retrotranscribed into cDNA, normalized to that of the Hprt gene, and expressed as time-fold variations relative to the values obtained from wild-type (WT) animals. Solid blue, WT; blue outline, PGC-1α : MCK-PGC-1α transgenic mouse; solid red, Surf1 −/− , constitutive Surf1 knockout mouse; red outline, Surf1 −/− /PGC-1α : Surf1 −/− /MCK-PGC-1α double mutant mouse. Error bars represent the standard deviation (SD). (Right panel) Western blot immunovisualization of skeletal muscle proteins of the different genotypes, listed as above. Densitometric analysis of each band, normalized against that of GAPDH, revealed the following variations, relative to WT. (1) PGC-1α, 4.3 in MCK-PGC-1α ; 0.5 in Surf1 −/− ; 2.2 in Surf1 −/− /PGC-1α . (2) COX1, 1.8 in MCK-PGC-1α ; 0.2 in Surf1 −/− ; 1.0 in Surf1 −/− /PGC-1α . (3) COX5a, 5.0 in MCK-PGC-1α ; ∼0.0 in Surf1 −/− ; 1.0 in Surf1 −/− /PGC-1α . (B) MtDNA analysis in different genotypes (three animals/genotype). Color code is as in (A). MtDNA is expressed as number of DNA molecules per cell ( Cree et al., 2008 ). Error bars represent SD. (C) MRC activities in the different genotypes (three animals/genotype), expressed as nmoles/min/mg of protein. Note that the activity of cII has been multiplied by 10 for visualization clarity. Color code is as in (A). CS, citrate synthase; CI-IV, MRC complexes I–IV. Error bars represent the standard deviation (SD). The asterisks represent the significance levels calculated by unpaired, Student's two-tailed t test: ∗ p

    Journal: Cell Metabolism

    Article Title: In Vivo Correction of COX Deficiency by Activation of the AMPK/PGC-1? Axis

    doi: 10.1016/j.cmet.2011.04.011

    Figure Lengend Snippet: Effects of PGC-1α Overexpression in Surf1 −/− Mice (A) Expression analysis of PGC-1α . (Left panel) The PGC-1α transcript from muscle of each mouse genotype (three animals/genotype) was retrotranscribed into cDNA, normalized to that of the Hprt gene, and expressed as time-fold variations relative to the values obtained from wild-type (WT) animals. Solid blue, WT; blue outline, PGC-1α : MCK-PGC-1α transgenic mouse; solid red, Surf1 −/− , constitutive Surf1 knockout mouse; red outline, Surf1 −/− /PGC-1α : Surf1 −/− /MCK-PGC-1α double mutant mouse. Error bars represent the standard deviation (SD). (Right panel) Western blot immunovisualization of skeletal muscle proteins of the different genotypes, listed as above. Densitometric analysis of each band, normalized against that of GAPDH, revealed the following variations, relative to WT. (1) PGC-1α, 4.3 in MCK-PGC-1α ; 0.5 in Surf1 −/− ; 2.2 in Surf1 −/− /PGC-1α . (2) COX1, 1.8 in MCK-PGC-1α ; 0.2 in Surf1 −/− ; 1.0 in Surf1 −/− /PGC-1α . (3) COX5a, 5.0 in MCK-PGC-1α ; ∼0.0 in Surf1 −/− ; 1.0 in Surf1 −/− /PGC-1α . (B) MtDNA analysis in different genotypes (three animals/genotype). Color code is as in (A). MtDNA is expressed as number of DNA molecules per cell ( Cree et al., 2008 ). Error bars represent SD. (C) MRC activities in the different genotypes (three animals/genotype), expressed as nmoles/min/mg of protein. Note that the activity of cII has been multiplied by 10 for visualization clarity. Color code is as in (A). CS, citrate synthase; CI-IV, MRC complexes I–IV. Error bars represent the standard deviation (SD). The asterisks represent the significance levels calculated by unpaired, Student's two-tailed t test: ∗ p

    Article Snippet: Anti-PGC-1α antibody was from Abcam; anti-AMPK and anti-AMPK-P antibodies were from Cell Signaling; anti-COI and -COX5a were from Invitrogen; anti-GAPDH was from Millipore.

    Techniques: Pyrolysis Gas Chromatography, Over Expression, Mouse Assay, Expressing, Transgenic Assay, Knock-Out, Mutagenesis, Standard Deviation, Western Blot, Activity Assay, Two Tailed Test

    RNA sequencing analysis indicates that PGC1α induces aberrant mitochondrial assembly and promotes cell cycle re-entry.

    Journal: JCI Insight

    Article Title: Increasing the level of peroxisome proliferator-activated receptor γ coactivator-1 α in podocytes results in collapsing glomerulopathy

    doi: 10.1172/jci.insight.92930

    Figure Lengend Snippet: RNA sequencing analysis indicates that PGC1α induces aberrant mitochondrial assembly and promotes cell cycle re-entry.

    Article Snippet: Immunofluorescence double staining was performed with antibodies against PGC1α, WT-1, and claudin 1 (ab15098, Abcam) in OCT-embedded frozen sections.

    Techniques: RNA Sequencing Assay

    PGC1α induces podocyte proliferation in vivo.

    Journal: JCI Insight

    Article Title: Increasing the level of peroxisome proliferator-activated receptor γ coactivator-1 α in podocytes results in collapsing glomerulopathy

    doi: 10.1172/jci.insight.92930

    Figure Lengend Snippet: PGC1α induces podocyte proliferation in vivo.

    Article Snippet: Immunofluorescence double staining was performed with antibodies against PGC1α, WT-1, and claudin 1 (ab15098, Abcam) in OCT-embedded frozen sections.

    Techniques: In Vivo

    Excessive PGC1α alters mitochondrial properties in vivo.

    Journal: JCI Insight

    Article Title: Increasing the level of peroxisome proliferator-activated receptor γ coactivator-1 α in podocytes results in collapsing glomerulopathy

    doi: 10.1172/jci.insight.92930

    Figure Lengend Snippet: Excessive PGC1α alters mitochondrial properties in vivo.

    Article Snippet: Immunofluorescence double staining was performed with antibodies against PGC1α, WT-1, and claudin 1 (ab15098, Abcam) in OCT-embedded frozen sections.

    Techniques: In Vivo

    Nefta-PGC1α mice develop collapsing glomerulosclerosis.

    Journal: JCI Insight

    Article Title: Increasing the level of peroxisome proliferator-activated receptor γ coactivator-1 α in podocytes results in collapsing glomerulopathy

    doi: 10.1172/jci.insight.92930

    Figure Lengend Snippet: Nefta-PGC1α mice develop collapsing glomerulosclerosis.

    Article Snippet: Immunofluorescence double staining was performed with antibodies against PGC1α, WT-1, and claudin 1 (ab15098, Abcam) in OCT-embedded frozen sections.

    Techniques: Mouse Assay

    Increasing PGC1α levels in podocytes leads to proteinuria and renal failure.

    Journal: JCI Insight

    Article Title: Increasing the level of peroxisome proliferator-activated receptor γ coactivator-1 α in podocytes results in collapsing glomerulopathy

    doi: 10.1172/jci.insight.92930

    Figure Lengend Snippet: Increasing PGC1α levels in podocytes leads to proteinuria and renal failure.

    Article Snippet: Immunofluorescence double staining was performed with antibodies against PGC1α, WT-1, and claudin 1 (ab15098, Abcam) in OCT-embedded frozen sections.

    Techniques:

    PGC1α increase mitochondrial biogenesis, mitochondrial fusion, and mitochondrial respiratory capacity in vitro.

    Journal: JCI Insight

    Article Title: Increasing the level of peroxisome proliferator-activated receptor γ coactivator-1 α in podocytes results in collapsing glomerulopathy

    doi: 10.1172/jci.insight.92930

    Figure Lengend Snippet: PGC1α increase mitochondrial biogenesis, mitochondrial fusion, and mitochondrial respiratory capacity in vitro.

    Article Snippet: Immunofluorescence double staining was performed with antibodies against PGC1α, WT-1, and claudin 1 (ab15098, Abcam) in OCT-embedded frozen sections.

    Techniques: In Vitro

    PGC1α alters mitochondrial fuel preference in podocytes.

    Journal: JCI Insight

    Article Title: Increasing the level of peroxisome proliferator-activated receptor γ coactivator-1 α in podocytes results in collapsing glomerulopathy

    doi: 10.1172/jci.insight.92930

    Figure Lengend Snippet: PGC1α alters mitochondrial fuel preference in podocytes.

    Article Snippet: Immunofluorescence double staining was performed with antibodies against PGC1α, WT-1, and claudin 1 (ab15098, Abcam) in OCT-embedded frozen sections.

    Techniques:

    PGC1α induces podocyte proliferation in vitro.

    Journal: JCI Insight

    Article Title: Increasing the level of peroxisome proliferator-activated receptor γ coactivator-1 α in podocytes results in collapsing glomerulopathy

    doi: 10.1172/jci.insight.92930

    Figure Lengend Snippet: PGC1α induces podocyte proliferation in vitro.

    Article Snippet: Immunofluorescence double staining was performed with antibodies against PGC1α, WT-1, and claudin 1 (ab15098, Abcam) in OCT-embedded frozen sections.

    Techniques: In Vitro

    Glomerular PGC1α expression and mitochondrial transcript abundance in diabetic glomeruli.

    Journal: JCI Insight

    Article Title: Increasing the level of peroxisome proliferator-activated receptor γ coactivator-1 α in podocytes results in collapsing glomerulopathy

    doi: 10.1172/jci.insight.92930

    Figure Lengend Snippet: Glomerular PGC1α expression and mitochondrial transcript abundance in diabetic glomeruli.

    Article Snippet: Immunofluorescence double staining was performed with antibodies against PGC1α, WT-1, and claudin 1 (ab15098, Abcam) in OCT-embedded frozen sections.

    Techniques: Expressing

    Loss of perlecan modifies the composition of myosin heavy chains by activating PGC1α. Detection by ( a ) SDS-PAGE-coupled silver staining and ( b ) relative composition of myosin heavy chain isoforms in the quadriceps of the WT-Tg and Hspg2 −/− -Tg mice. Soleus and plantaris represent markers for type I and II fibers, respectively. The relative intensity of the bands was quantified using ImageJ software. ( c–e ) Protein expression levels of ( c ) translocase of outer membrane 20 (TOM20), ( d ) translocase of inner membrane 23 (TIM23), and ( e ) peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) in the quadriceps of the WT-Tg and Hspg2 −/− -Tg mice. ( f ) Representative images of proteins extracted from quadriceps and stained with Ponceau S (Ponc) after SDS-PAGE. The relative intensities of the respective bands detected by western blotting using the specific antibody to the Ponc-stained patterns were quantified using ImageJ software. Data points and error bars represent the mean ± S.D. (n = 5 in a and b ; n = 5–6 in c – e ). Data were analyzed by two-way ANOVA with Sidak’s multiple comparison ( b ) and unpaired t- test ( c – e ). * p

    Journal: Scientific Reports

    Article Title: Perlecan, a heparan sulfate proteoglycan, regulates systemic metabolism with dynamic changes in adipose tissue and skeletal muscle

    doi: 10.1038/s41598-018-25635-x

    Figure Lengend Snippet: Loss of perlecan modifies the composition of myosin heavy chains by activating PGC1α. Detection by ( a ) SDS-PAGE-coupled silver staining and ( b ) relative composition of myosin heavy chain isoforms in the quadriceps of the WT-Tg and Hspg2 −/− -Tg mice. Soleus and plantaris represent markers for type I and II fibers, respectively. The relative intensity of the bands was quantified using ImageJ software. ( c–e ) Protein expression levels of ( c ) translocase of outer membrane 20 (TOM20), ( d ) translocase of inner membrane 23 (TIM23), and ( e ) peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) in the quadriceps of the WT-Tg and Hspg2 −/− -Tg mice. ( f ) Representative images of proteins extracted from quadriceps and stained with Ponceau S (Ponc) after SDS-PAGE. The relative intensities of the respective bands detected by western blotting using the specific antibody to the Ponc-stained patterns were quantified using ImageJ software. Data points and error bars represent the mean ± S.D. (n = 5 in a and b ; n = 5–6 in c – e ). Data were analyzed by two-way ANOVA with Sidak’s multiple comparison ( b ) and unpaired t- test ( c – e ). * p

    Article Snippet: The primary antibodies used were: anti-TIM23 antibody (1:1,000 dilution, #611222, BD Biosciences), anti-TOM20 (FL-145) antibody (1:500, sc-11415, Santa Cruz Biotechnology), and anti-PGC1α antibody (1:1,000, ab54481, abcam).

    Techniques: SDS Page, Silver Staining, Mouse Assay, Software, Expressing, Staining, Western Blot

    Effects of flavan-3-ols on expression of mitochondrial biogenesis-related protein in insulin-resistant 3T3-L1 adipocytes. RT-PCR analysis of mRNA levels of, ( a ) PGC-1α, ( b ) NRF1, and ( c ) Tfam. ( d ) Western blot analysis of expression of SirT1 and PGC-1α. Fully differentiated 3T3-L1 adipocytes were treated with Dex for 48 h in the presence or absence of flavan-3-ols or rosiglitazone. Con, normal differentiated 3T3-L1 adipocytes; Mod, Dex alone treated differentiated 3T3-L1 adipocytes; Rosi, rosiglitazone; C, catechin; EC, epicatechin; PB7, procyanidin B7; PB6, procyanidin B6; PB3, procyanidin B3; PB1, procyanidin B1. Values are shown as Mean ± SD of three independent triplicate experiments ( n = 3). # p

    Journal: International Journal of Molecular Sciences

    Article Title: Proanthocyanidins Ameliorated Deficits of Lipid Metabolism in Type 2 Diabetes Mellitus Via Inhibiting Adipogenesis and Improving Mitochondrial Function

    doi: 10.3390/ijms21062029

    Figure Lengend Snippet: Effects of flavan-3-ols on expression of mitochondrial biogenesis-related protein in insulin-resistant 3T3-L1 adipocytes. RT-PCR analysis of mRNA levels of, ( a ) PGC-1α, ( b ) NRF1, and ( c ) Tfam. ( d ) Western blot analysis of expression of SirT1 and PGC-1α. Fully differentiated 3T3-L1 adipocytes were treated with Dex for 48 h in the presence or absence of flavan-3-ols or rosiglitazone. Con, normal differentiated 3T3-L1 adipocytes; Mod, Dex alone treated differentiated 3T3-L1 adipocytes; Rosi, rosiglitazone; C, catechin; EC, epicatechin; PB7, procyanidin B7; PB6, procyanidin B6; PB3, procyanidin B3; PB1, procyanidin B1. Values are shown as Mean ± SD of three independent triplicate experiments ( n = 3). # p

    Article Snippet: Antibodies of β-actin, FAS, ACC, ATGL, HSL, CPT1A, p-AMPK, AMPK, PPARγ, C/EBPα, FABP4, AKT, p-AKT, Mfn1, Mfn2, Drp1, SirT1 and PGC-1α were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Pyrolysis Gas Chromatography, Western Blot

    GH administration upregulated hepatic SIRT1 expression in alcohol-fed mice . (A) Hepatic mRNA expression of SIRT1. (B) SIRT1 protein levels. (C) Relative PGC1α acetylation. Hepatic nuclear SIRT1 protein levels were determined using an anti-SIRT1 antibody. A nonspecific nuclear protein band was used to confirm equal loading and to normalize the data. PGC1α was immunoprecipitated from liver extracts and immunoblotted with either an anti-acetylated lysine (Ac-Lys) antibody to determine the extent of PGC1α acetylation or with an anti-PGC1α antibody to determine the total amount of PGC1α. C: control group; GC: GH1-treated control group; A: alcohol group; GA: GH1-treated alcohol group; PI: pair-fed group I; PII: pair-fed group II. AOX: acyl-CoA oxidase; CPT1a: carnitine palmitoyltransferase 1a; MCDA: medium chain acyl-Co-A dehydrogenase; PGC1α: PPARα coactivator; PPARγ: peroxisome proliferator activated receptor-γ; SIRT1: sirtuin 1. n = 6 mice per group. Means without a common letter differ at P

    Journal: Journal of Translational Medicine

    Article Title: Exploring the molecular mechanisms underlying the potentiation of exogenous growth hormone on alcohol-induced fatty liver diseases in mice

    doi: 10.1186/1479-5876-8-120

    Figure Lengend Snippet: GH administration upregulated hepatic SIRT1 expression in alcohol-fed mice . (A) Hepatic mRNA expression of SIRT1. (B) SIRT1 protein levels. (C) Relative PGC1α acetylation. Hepatic nuclear SIRT1 protein levels were determined using an anti-SIRT1 antibody. A nonspecific nuclear protein band was used to confirm equal loading and to normalize the data. PGC1α was immunoprecipitated from liver extracts and immunoblotted with either an anti-acetylated lysine (Ac-Lys) antibody to determine the extent of PGC1α acetylation or with an anti-PGC1α antibody to determine the total amount of PGC1α. C: control group; GC: GH1-treated control group; A: alcohol group; GA: GH1-treated alcohol group; PI: pair-fed group I; PII: pair-fed group II. AOX: acyl-CoA oxidase; CPT1a: carnitine palmitoyltransferase 1a; MCDA: medium chain acyl-Co-A dehydrogenase; PGC1α: PPARα coactivator; PPARγ: peroxisome proliferator activated receptor-γ; SIRT1: sirtuin 1. n = 6 mice per group. Means without a common letter differ at P

    Article Snippet: Total AMPKα, phospho-AMPKα (p-AMPKα), phospho-ACC (p-ACC) and PGC1α were visualized using primary antibodies from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Expressing, Mouse Assay, Immunoprecipitation

    Markers of mitochondrial function. ( A ) Mitochondrial protein content assessed by Western blot and ( B ) mitochondrial maximal enzyme activity in skeletal muscle of lean ( n = 9) and obese ( n = 9) men prior to and following 12-wk endurance training. ( A ) PGC-1α, peroxisome proliferator-activated receptor-γ coactivator-1α; CS, citrate synthase; COX, cytochrome c oxidase - subunits II and IV. Results were normalized to β-actin protein content. * P ≤0.04 pre- vs. post-training (main effect). ( B ) CS, citrate synthase; COX, cytochrome c oxidase; SCHAD, short-chain β-hydroxyacyl-CoA dehydrogenase. * P ≤0.03 pre- vs. post-training (main effect). ( C ) Mitochondrial DNA (mtDNA) copy number determined by real-time quantitative PCR using a TaqMan probe against NADH dehydrogenase 4 (ND4) and β-globin. mtDNA copy number was calculated as the ratio of ND4 to β-globin in skeletal muscle of lean ( n = 3) and obese ( n = 5) men prior to and following 12-wk endurance training. * P = 0.10 pre- vs. post-training (main effect).

    Journal: PLoS ONE

    Article Title: Markers of Skeletal Muscle Mitochondrial Function and Lipid Accumulation Are Moderately Associated with the Homeostasis Model Assessment Index of Insulin Resistance in Obese Men

    doi: 10.1371/journal.pone.0066322

    Figure Lengend Snippet: Markers of mitochondrial function. ( A ) Mitochondrial protein content assessed by Western blot and ( B ) mitochondrial maximal enzyme activity in skeletal muscle of lean ( n = 9) and obese ( n = 9) men prior to and following 12-wk endurance training. ( A ) PGC-1α, peroxisome proliferator-activated receptor-γ coactivator-1α; CS, citrate synthase; COX, cytochrome c oxidase - subunits II and IV. Results were normalized to β-actin protein content. * P ≤0.04 pre- vs. post-training (main effect). ( B ) CS, citrate synthase; COX, cytochrome c oxidase; SCHAD, short-chain β-hydroxyacyl-CoA dehydrogenase. * P ≤0.03 pre- vs. post-training (main effect). ( C ) Mitochondrial DNA (mtDNA) copy number determined by real-time quantitative PCR using a TaqMan probe against NADH dehydrogenase 4 (ND4) and β-globin. mtDNA copy number was calculated as the ratio of ND4 to β-globin in skeletal muscle of lean ( n = 3) and obese ( n = 5) men prior to and following 12-wk endurance training. * P = 0.10 pre- vs. post-training (main effect).

    Article Snippet: Proteins were resolved on either 7.5, 10 or 12.5% SDS-PAGE gels, transferred onto Hybond® ECL nitrocellulose membranes (Amersham), and immunoblotted using the following commercially available primary antibodies: anti-COX subunit II (cytochrome c oxidase - subunit II, MS405) and anti-COX subunit IV (cytochrome c oxidase - subunit IV, MS408) were purchased from MitoSciences; anti-GLUT4 from Chemicon (ab1346); anti-phospho-Akt (Ser473 , 4060) and anti-PGC-1α (peroxisome proliferator-activated receptor-γ coactivator-1α, 4187) from Cell Signaling Technology.

    Techniques: Western Blot, Activity Assay, Pyrolysis Gas Chromatography, Real-time Polymerase Chain Reaction

    AgNPs disrupted mitochondrial and thermogenic functions of beige adipocytes . (A) Experimental schematics of AgNP administration on differentiated adipocytes. (B) Basal oxygen consumption in primary beige adipocytes. (C) Thermogenic and mitochondrial gene programs in C3H10T1/2 adipocytes under different Ag20NP doses. (D) AgNPs blocked forskolin (FSK)-induced elevation in the thermogenic (left) and mitochondrial (right) gene programs in C3H10T1/2 adipocytes. (E) AgNPs blocked forskolin (FSK)-induced elevation in PGC1α and UCP1 protein levels. (F) Ag20NP treatment inhibited thermogenic and mitochondrial gene programs in primary beige adipocytes. Data are presented as mean ± SEM and *P

    Journal: Molecular Metabolism

    Article Title: Silver nanoparticles inhibit beige fat function and promote adiposity

    doi: 10.1016/j.molmet.2019.01.005

    Figure Lengend Snippet: AgNPs disrupted mitochondrial and thermogenic functions of beige adipocytes . (A) Experimental schematics of AgNP administration on differentiated adipocytes. (B) Basal oxygen consumption in primary beige adipocytes. (C) Thermogenic and mitochondrial gene programs in C3H10T1/2 adipocytes under different Ag20NP doses. (D) AgNPs blocked forskolin (FSK)-induced elevation in the thermogenic (left) and mitochondrial (right) gene programs in C3H10T1/2 adipocytes. (E) AgNPs blocked forskolin (FSK)-induced elevation in PGC1α and UCP1 protein levels. (F) Ag20NP treatment inhibited thermogenic and mitochondrial gene programs in primary beige adipocytes. Data are presented as mean ± SEM and *P

    Article Snippet: After blocking with 5% skimmed milk, the membrane was incubated overnight at 4 °C with primary antibodies, including anti-UCP1 (Abcam, ab10983), anti-PGC1α (Santa Biotechnology, sc-517380) and anti-β-actin (Santa Biotechnology, sc-47778), antibodies against MAPKs were purchased from Cell Signaling Technology including p-p38 (4511S), p38α (2371S), p-JNK(4671S), JNK(9252S), p-ERK (9106S) and ERK (4695S).

    Techniques:

    Impact of AgNPs on beige adipocytes and mouse physiology . After AgNPs exposure, nanoparticles enter the cytoplasm and reside in lysosomes, releasing ROS into the lysosomal acidic environment. The elevated, cellular ROS levels cause oxidative stress and increase ERK phosphorylation, which, in turn, suppress thermogenic and mitochondrial gene programs including PGC1α and UCP1. AgNPs exposure caused mitochondrial dysfunction leads to decreased energy expenditure and increased adiposity in mice.

    Journal: Molecular Metabolism

    Article Title: Silver nanoparticles inhibit beige fat function and promote adiposity

    doi: 10.1016/j.molmet.2019.01.005

    Figure Lengend Snippet: Impact of AgNPs on beige adipocytes and mouse physiology . After AgNPs exposure, nanoparticles enter the cytoplasm and reside in lysosomes, releasing ROS into the lysosomal acidic environment. The elevated, cellular ROS levels cause oxidative stress and increase ERK phosphorylation, which, in turn, suppress thermogenic and mitochondrial gene programs including PGC1α and UCP1. AgNPs exposure caused mitochondrial dysfunction leads to decreased energy expenditure and increased adiposity in mice.

    Article Snippet: After blocking with 5% skimmed milk, the membrane was incubated overnight at 4 °C with primary antibodies, including anti-UCP1 (Abcam, ab10983), anti-PGC1α (Santa Biotechnology, sc-517380) and anti-β-actin (Santa Biotechnology, sc-47778), antibodies against MAPKs were purchased from Cell Signaling Technology including p-p38 (4511S), p38α (2371S), p-JNK(4671S), JNK(9252S), p-ERK (9106S) and ERK (4695S).

    Techniques: Mouse Assay

    Mitochondrial ultrastructural abnormalities in hearts of adult PGC-1αβ −/− mice. Representative electron micrographs of sections taken from the left ventricular papillary muscle from 16-week-old PGC-1α −/− (α −/− ) and PGC-1αβ −/− (αβ −/− ) mice at two different magnifications. Arrows indicate the structurally abnormal mitochondria with “stacked” cristae ( white arrows ). Scale bar for each magnification is shown at the bottom right corner : A , 1 μm; B , 5 μm.

    Journal: The Journal of Biological Chemistry

    Article Title: A Role for Peroxisome Proliferator-activated Receptor γ Coactivator 1 (PGC-1) in the Regulation of Cardiac Mitochondrial Phospholipid Biosynthesis *A Role for Peroxisome Proliferator-activated Receptor γ Coactivator 1 (PGC-1) in the Regulation of Cardiac Mitochondrial Phospholipid Biosynthesis * ♦

    doi: 10.1074/jbc.M113.523654

    Figure Lengend Snippet: Mitochondrial ultrastructural abnormalities in hearts of adult PGC-1αβ −/− mice. Representative electron micrographs of sections taken from the left ventricular papillary muscle from 16-week-old PGC-1α −/− (α −/− ) and PGC-1αβ −/− (αβ −/− ) mice at two different magnifications. Arrows indicate the structurally abnormal mitochondria with “stacked” cristae ( white arrows ). Scale bar for each magnification is shown at the bottom right corner : A , 1 μm; B , 5 μm.

    Article Snippet: Sheared protein-DNA complexes were immunoprecipitated by using anti-ERRα , anti-PGC-1α ( , ), or rabbit IgG control (Sigma).

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay

    Altered expression of phospholipid biosynthesis pathway genes in adult PGC-1αβ −/− mice. A, schematic representation of de novo PL biosynthesis and remodeling pathways in mammalian cells. B, results of qRT-PCR analysis of RNA extracted from hearts of 12-week-old wild-type (αβ +/+ ), PGC-1α −/− (α −/− ), PGC-1β −/− (β −/− ), and PGC-1αβ −/− (αβ −/− ) mice for the following: CDP-DAG synthase ( Cds ); phosphatidylglycerophosphate synthase 1 ( Pgs1 ); protein-tyrosine phosphatase, mitochondrial 1 ( Ptpmt1 ); cardiolipin synthase ( Crls ); tafazzin ( Taz ); lipin 1 ( Lpin1 ); phosphatidic acid phosphatase type 2C ( Chptc ); choline phosphotransferase 1 ( Chpt1 ); choline/ethanolaminephosphotransferase 1 ( Cept1 ); phosphate cytidylyltransferase 1; choline ( Pcyt1 ); and phosphate cytidylyltransferase 2, ethanolamine ( Pcyt2 ). Bars represent mean ± S.E. *, p

    Journal: The Journal of Biological Chemistry

    Article Title: A Role for Peroxisome Proliferator-activated Receptor γ Coactivator 1 (PGC-1) in the Regulation of Cardiac Mitochondrial Phospholipid Biosynthesis *A Role for Peroxisome Proliferator-activated Receptor γ Coactivator 1 (PGC-1) in the Regulation of Cardiac Mitochondrial Phospholipid Biosynthesis * ♦

    doi: 10.1074/jbc.M113.523654

    Figure Lengend Snippet: Altered expression of phospholipid biosynthesis pathway genes in adult PGC-1αβ −/− mice. A, schematic representation of de novo PL biosynthesis and remodeling pathways in mammalian cells. B, results of qRT-PCR analysis of RNA extracted from hearts of 12-week-old wild-type (αβ +/+ ), PGC-1α −/− (α −/− ), PGC-1β −/− (β −/− ), and PGC-1αβ −/− (αβ −/− ) mice for the following: CDP-DAG synthase ( Cds ); phosphatidylglycerophosphate synthase 1 ( Pgs1 ); protein-tyrosine phosphatase, mitochondrial 1 ( Ptpmt1 ); cardiolipin synthase ( Crls ); tafazzin ( Taz ); lipin 1 ( Lpin1 ); phosphatidic acid phosphatase type 2C ( Chptc ); choline phosphotransferase 1 ( Chpt1 ); choline/ethanolaminephosphotransferase 1 ( Cept1 ); phosphate cytidylyltransferase 1; choline ( Pcyt1 ); and phosphate cytidylyltransferase 2, ethanolamine ( Pcyt2 ). Bars represent mean ± S.E. *, p

    Article Snippet: Sheared protein-DNA complexes were immunoprecipitated by using anti-ERRα , anti-PGC-1α ( , ), or rabbit IgG control (Sigma).

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

    Transcriptional activation of the Cds1 gene by PGC-1/ERR. A, transient transfections were performed in C2C12 myoblasts (differentiated into myotubes) using a mouse Cds1 promoter reporter construct, mcds1.luc.2.39, containing a putative ERR-binding site at −738 and +1294, as denoted in B , cotransfected with pcDNA3.1-PGC-1α construct (+) compared with vector backbone alone (−). The bars represent mean ± S.E. promoter-reporter activity shown as relative light units ( RLU ) normalized to the condition transfected with vector backbone alone. C, results of transient transfection performed with mouse Cds1 reporter mcds1.luc.2.39 and truncation mutants of mcds1.luc.1.97, 1.15, or 0.73 with the pcDNA3.1-PGC-1α construct in C2C12 myoblasts (differentiated into myotubes). D, C2C12 myoblasts were transfected with mouse Cds1 reporter mcds1.luc.2.39 in the presence of pcDNA3.1-PGC-1α or pCATCH-PGC-1β and expression vector of ERRα, ERRβ, or ERRγ. *, p

    Journal: The Journal of Biological Chemistry

    Article Title: A Role for Peroxisome Proliferator-activated Receptor γ Coactivator 1 (PGC-1) in the Regulation of Cardiac Mitochondrial Phospholipid Biosynthesis *A Role for Peroxisome Proliferator-activated Receptor γ Coactivator 1 (PGC-1) in the Regulation of Cardiac Mitochondrial Phospholipid Biosynthesis * ♦

    doi: 10.1074/jbc.M113.523654

    Figure Lengend Snippet: Transcriptional activation of the Cds1 gene by PGC-1/ERR. A, transient transfections were performed in C2C12 myoblasts (differentiated into myotubes) using a mouse Cds1 promoter reporter construct, mcds1.luc.2.39, containing a putative ERR-binding site at −738 and +1294, as denoted in B , cotransfected with pcDNA3.1-PGC-1α construct (+) compared with vector backbone alone (−). The bars represent mean ± S.E. promoter-reporter activity shown as relative light units ( RLU ) normalized to the condition transfected with vector backbone alone. C, results of transient transfection performed with mouse Cds1 reporter mcds1.luc.2.39 and truncation mutants of mcds1.luc.1.97, 1.15, or 0.73 with the pcDNA3.1-PGC-1α construct in C2C12 myoblasts (differentiated into myotubes). D, C2C12 myoblasts were transfected with mouse Cds1 reporter mcds1.luc.2.39 in the presence of pcDNA3.1-PGC-1α or pCATCH-PGC-1β and expression vector of ERRα, ERRβ, or ERRγ. *, p

    Article Snippet: Sheared protein-DNA complexes were immunoprecipitated by using anti-ERRα , anti-PGC-1α ( , ), or rabbit IgG control (Sigma).

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

    Liver PGC-1α overexpression produces elevated liver FAO. After adenoviral transduction, FAO was determined in liver homogenates. A–C : AdvPGC1a-transduced animals had higher complete FAO to CO 2 , with no difference in total FAO, which results

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: PGC-1? overexpression results in increased hepatic fatty acid oxidation with reduced triacylglycerol accumulation and secretion

    doi: 10.1152/ajpgi.00169.2012

    Figure Lengend Snippet: Liver PGC-1α overexpression produces elevated liver FAO. After adenoviral transduction, FAO was determined in liver homogenates. A–C : AdvPGC1a-transduced animals had higher complete FAO to CO 2 , with no difference in total FAO, which results

    Article Snippet: PGC-1α antibody was purchased from Calbiochem (EMD Chemicals, Gibbstown, NJ), apoB antibody from Abcam (Cambridge, MA), MTTP antibodies from Santa Cruz Biotechnology (Santa Cruz, CA), mitochondrial transcription factor A (mtTFA) antibody from Novus Biologicals (Littleton, CO), and oxidative phosphorylation complex I–V antibody cocktail from MitoScience (Eugene, OR).

    Techniques: Pyrolysis Gas Chromatography, Over Expression, Transduction

    Adenoviral PGC-1α (AdvPGC1a) transduction increases hepatic PGC-1α expression and citrate synthase activity. Liver PGC-1α expression following adenoviral transduction was determined by RT-PCR and Western blot analysis. Citrate

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: PGC-1? overexpression results in increased hepatic fatty acid oxidation with reduced triacylglycerol accumulation and secretion

    doi: 10.1152/ajpgi.00169.2012

    Figure Lengend Snippet: Adenoviral PGC-1α (AdvPGC1a) transduction increases hepatic PGC-1α expression and citrate synthase activity. Liver PGC-1α expression following adenoviral transduction was determined by RT-PCR and Western blot analysis. Citrate

    Article Snippet: PGC-1α antibody was purchased from Calbiochem (EMD Chemicals, Gibbstown, NJ), apoB antibody from Abcam (Cambridge, MA), MTTP antibodies from Santa Cruz Biotechnology (Santa Cruz, CA), mitochondrial transcription factor A (mtTFA) antibody from Novus Biologicals (Littleton, CO), and oxidative phosphorylation complex I–V antibody cocktail from MitoScience (Eugene, OR).

    Techniques: Pyrolysis Gas Chromatography, Transduction, Expressing, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot

    Adenoviral overexpression of PGC-1α increases primary hepatocyte fatty acid oxidation (FAO). FAO rates of 14 C-radiolabeled palmitate-oleate (50 μM palmitate-50 μM oleate) by primary hepatocytes were determined in the presence and

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: PGC-1? overexpression results in increased hepatic fatty acid oxidation with reduced triacylglycerol accumulation and secretion

    doi: 10.1152/ajpgi.00169.2012

    Figure Lengend Snippet: Adenoviral overexpression of PGC-1α increases primary hepatocyte fatty acid oxidation (FAO). FAO rates of 14 C-radiolabeled palmitate-oleate (50 μM palmitate-50 μM oleate) by primary hepatocytes were determined in the presence and

    Article Snippet: PGC-1α antibody was purchased from Calbiochem (EMD Chemicals, Gibbstown, NJ), apoB antibody from Abcam (Cambridge, MA), MTTP antibodies from Santa Cruz Biotechnology (Santa Cruz, CA), mitochondrial transcription factor A (mtTFA) antibody from Novus Biologicals (Littleton, CO), and oxidative phosphorylation complex I–V antibody cocktail from MitoScience (Eugene, OR).

    Techniques: Over Expression, Pyrolysis Gas Chromatography

    Adenoviral overexpression of PGC-1α in liver reduces plasma and tissue TAG accumulation. Liver TAG accumulation and plasma TAG levels were determined to assess hepatic lipid handling. A and B : AdvPGC1a-transduced rats demonstrated decreased hepatic

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: PGC-1? overexpression results in increased hepatic fatty acid oxidation with reduced triacylglycerol accumulation and secretion

    doi: 10.1152/ajpgi.00169.2012

    Figure Lengend Snippet: Adenoviral overexpression of PGC-1α in liver reduces plasma and tissue TAG accumulation. Liver TAG accumulation and plasma TAG levels were determined to assess hepatic lipid handling. A and B : AdvPGC1a-transduced rats demonstrated decreased hepatic

    Article Snippet: PGC-1α antibody was purchased from Calbiochem (EMD Chemicals, Gibbstown, NJ), apoB antibody from Abcam (Cambridge, MA), MTTP antibodies from Santa Cruz Biotechnology (Santa Cruz, CA), mitochondrial transcription factor A (mtTFA) antibody from Novus Biologicals (Littleton, CO), and oxidative phosphorylation complex I–V antibody cocktail from MitoScience (Eugene, OR).

    Techniques: Over Expression, Pyrolysis Gas Chromatography

    Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) overexpression increases markers of primary hepatocyte mitochondrial biogenesis and content. Hepatocyte PGC-1α expression following adenoviral transduction

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: PGC-1? overexpression results in increased hepatic fatty acid oxidation with reduced triacylglycerol accumulation and secretion

    doi: 10.1152/ajpgi.00169.2012

    Figure Lengend Snippet: Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) overexpression increases markers of primary hepatocyte mitochondrial biogenesis and content. Hepatocyte PGC-1α expression following adenoviral transduction

    Article Snippet: PGC-1α antibody was purchased from Calbiochem (EMD Chemicals, Gibbstown, NJ), apoB antibody from Abcam (Cambridge, MA), MTTP antibodies from Santa Cruz Biotechnology (Santa Cruz, CA), mitochondrial transcription factor A (mtTFA) antibody from Novus Biologicals (Littleton, CO), and oxidative phosphorylation complex I–V antibody cocktail from MitoScience (Eugene, OR).

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

    Decreased triacylglycerol (TAG) accumulation and secretion in PGC-1α-overexpressing primary hepatocytes. TAG accumulation and secretion in primary hepatocytes were determined following overnight treatment with control or free fatty acids (FFA;

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: PGC-1? overexpression results in increased hepatic fatty acid oxidation with reduced triacylglycerol accumulation and secretion

    doi: 10.1152/ajpgi.00169.2012

    Figure Lengend Snippet: Decreased triacylglycerol (TAG) accumulation and secretion in PGC-1α-overexpressing primary hepatocytes. TAG accumulation and secretion in primary hepatocytes were determined following overnight treatment with control or free fatty acids (FFA;

    Article Snippet: PGC-1α antibody was purchased from Calbiochem (EMD Chemicals, Gibbstown, NJ), apoB antibody from Abcam (Cambridge, MA), MTTP antibodies from Santa Cruz Biotechnology (Santa Cruz, CA), mitochondrial transcription factor A (mtTFA) antibody from Novus Biologicals (Littleton, CO), and oxidative phosphorylation complex I–V antibody cocktail from MitoScience (Eugene, OR).

    Techniques: Pyrolysis Gas Chromatography

    Decreased expression of apolipoprotein B (apoB) and microsomal triglyceride transfer protein (MTTP) in PGC-1α-overexpressing primary hepatocytes. Expression of genes involved in hepatocyte lipoprotein assembly was determined by RT-PCR and Western

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: PGC-1? overexpression results in increased hepatic fatty acid oxidation with reduced triacylglycerol accumulation and secretion

    doi: 10.1152/ajpgi.00169.2012

    Figure Lengend Snippet: Decreased expression of apolipoprotein B (apoB) and microsomal triglyceride transfer protein (MTTP) in PGC-1α-overexpressing primary hepatocytes. Expression of genes involved in hepatocyte lipoprotein assembly was determined by RT-PCR and Western

    Article Snippet: PGC-1α antibody was purchased from Calbiochem (EMD Chemicals, Gibbstown, NJ), apoB antibody from Abcam (Cambridge, MA), MTTP antibodies from Santa Cruz Biotechnology (Santa Cruz, CA), mitochondrial transcription factor A (mtTFA) antibody from Novus Biologicals (Littleton, CO), and oxidative phosphorylation complex I–V antibody cocktail from MitoScience (Eugene, OR).

    Techniques: Expressing, Pyrolysis Gas Chromatography, Reverse Transcription Polymerase Chain Reaction, Western Blot

    Liver PGC-1α overexpression produces elevated mitochondrial FAO and tricarboxylic acid (TCA) cycle flux. After adenoviral transduction, FAO and pyruvate oxidation were determined in isolated liver mitochondria. A–C : in isolated liver mitochondria,

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: PGC-1? overexpression results in increased hepatic fatty acid oxidation with reduced triacylglycerol accumulation and secretion

    doi: 10.1152/ajpgi.00169.2012

    Figure Lengend Snippet: Liver PGC-1α overexpression produces elevated mitochondrial FAO and tricarboxylic acid (TCA) cycle flux. After adenoviral transduction, FAO and pyruvate oxidation were determined in isolated liver mitochondria. A–C : in isolated liver mitochondria,

    Article Snippet: PGC-1α antibody was purchased from Calbiochem (EMD Chemicals, Gibbstown, NJ), apoB antibody from Abcam (Cambridge, MA), MTTP antibodies from Santa Cruz Biotechnology (Santa Cruz, CA), mitochondrial transcription factor A (mtTFA) antibody from Novus Biologicals (Littleton, CO), and oxidative phosphorylation complex I–V antibody cocktail from MitoScience (Eugene, OR).

    Techniques: Pyrolysis Gas Chromatography, Over Expression, Transduction, Isolation

    Knockdown of Irx3 repressed the differentiation of SVFs toward beige adipocytes in vitro. (A–B) (A) Relative mRNA and (B) protein levels of Irx3 in SVFs from IWAT were efficiently knocked down after a 3-day infection of mouse Irx3 lentiviral shRNA ( n = 3 for each group). (C–D) (C) Oil Red O staining and (D) TG concentration of the mature beige adipocytes under eight-day differentiation ( n = 6 for each group). (E) Relative mRNA expression of Ucp1 , Cidea , Pgc-1α , C/ebpβ , and Prdm16 at different time points during beige adipocyte differentiation from IWAT SVFs ( n = 3–4 for different groups). (F–G) (F) Relative mRNA ( n = 4) and (G) protein expression of brown adipocyte marker genes of the induced beige adipocytes from mouse IWAT SVTs for eight days with the infection of mouse Irx3 lentiviral shRNA, with or without CL-316,243 activation. (H) Relative mRNA expression of adipocyte differentiation-related genes in beige adipocytes ( n = 4). (I) Relative mRNA expression of the indicated genes in the induced brown adipocytes from mouse BAT SVFs for eight days, with the infection of mouse Irx3 lentiviral shRNA ( n = 4). (J) Relative mRNA levels of IRX3 in SVFs from human sWAT after a 3-day infection of human IRX3 lentiviral shRNA (left), and the mRNA levels of brown adipocyte marker genes in the SVFs under 2-day differentiation ( n = 4). (K) OCR measurement of the beige adipocytes under 5-day differentiation, with the infection of mouse Irx3 lentiviral shRNA ( n = 5). (L–N) (L) Ucp1 promoter-luciferase reporter activity was measured in HEK293T cells transfected with pEGFP-C1 vector or IRX3 for 48 h. (M) Schematic diagram of the mutant mouse Ucp1 promoter deleting ACATGTGT among the − 3470 to − 3463 bp region proximal to the TSS of the Ucp1 gene. (N) Transcriptional activity of wild-type or mutant Ucp1 promoter was measured with IRX3 overexpression. (O) qPCR of Ucp1 promoter binding region which was recruited by Irx3 antibody in 8-day induced beige adipocytes, as analyzed by ChIP. Fold enrichment of Ucp1 promoter was given ( n = 3). For the measurement of luciferase activity, cells were seeded on 24-well plate and transfected with 800 ng pEGFP-C1 vector or IRX3, 200 ng Ucp1 promoter construct, and 1 ng pRL-SV40, followed by the harvest for luciferase activity assessment using a dual-luciferase reporter assay system (Promega). Luciferase activity was corrected for Renilla luciferase activity ( n = 3–4 for each group). For qPCR data, mRNA expression was normalized to 36b4 for mouse genes and βACTIN for human genes. SVF cells were isolated from IWAT and BAT of C57BL/6J mice or human sWAT wherever mentioned, and IRX3 shRNA were introduced to cells 24 h after seeding. Data were presented as mean ± s.e.m. * P

    Journal: EBioMedicine

    Article Title: IRX3 Promotes the Browning of White Adipocytes and Its Rare Variants are Associated with Human Obesity Risk

    doi: 10.1016/j.ebiom.2017.09.010

    Figure Lengend Snippet: Knockdown of Irx3 repressed the differentiation of SVFs toward beige adipocytes in vitro. (A–B) (A) Relative mRNA and (B) protein levels of Irx3 in SVFs from IWAT were efficiently knocked down after a 3-day infection of mouse Irx3 lentiviral shRNA ( n = 3 for each group). (C–D) (C) Oil Red O staining and (D) TG concentration of the mature beige adipocytes under eight-day differentiation ( n = 6 for each group). (E) Relative mRNA expression of Ucp1 , Cidea , Pgc-1α , C/ebpβ , and Prdm16 at different time points during beige adipocyte differentiation from IWAT SVFs ( n = 3–4 for different groups). (F–G) (F) Relative mRNA ( n = 4) and (G) protein expression of brown adipocyte marker genes of the induced beige adipocytes from mouse IWAT SVTs for eight days with the infection of mouse Irx3 lentiviral shRNA, with or without CL-316,243 activation. (H) Relative mRNA expression of adipocyte differentiation-related genes in beige adipocytes ( n = 4). (I) Relative mRNA expression of the indicated genes in the induced brown adipocytes from mouse BAT SVFs for eight days, with the infection of mouse Irx3 lentiviral shRNA ( n = 4). (J) Relative mRNA levels of IRX3 in SVFs from human sWAT after a 3-day infection of human IRX3 lentiviral shRNA (left), and the mRNA levels of brown adipocyte marker genes in the SVFs under 2-day differentiation ( n = 4). (K) OCR measurement of the beige adipocytes under 5-day differentiation, with the infection of mouse Irx3 lentiviral shRNA ( n = 5). (L–N) (L) Ucp1 promoter-luciferase reporter activity was measured in HEK293T cells transfected with pEGFP-C1 vector or IRX3 for 48 h. (M) Schematic diagram of the mutant mouse Ucp1 promoter deleting ACATGTGT among the − 3470 to − 3463 bp region proximal to the TSS of the Ucp1 gene. (N) Transcriptional activity of wild-type or mutant Ucp1 promoter was measured with IRX3 overexpression. (O) qPCR of Ucp1 promoter binding region which was recruited by Irx3 antibody in 8-day induced beige adipocytes, as analyzed by ChIP. Fold enrichment of Ucp1 promoter was given ( n = 3). For the measurement of luciferase activity, cells were seeded on 24-well plate and transfected with 800 ng pEGFP-C1 vector or IRX3, 200 ng Ucp1 promoter construct, and 1 ng pRL-SV40, followed by the harvest for luciferase activity assessment using a dual-luciferase reporter assay system (Promega). Luciferase activity was corrected for Renilla luciferase activity ( n = 3–4 for each group). For qPCR data, mRNA expression was normalized to 36b4 for mouse genes and βACTIN for human genes. SVF cells were isolated from IWAT and BAT of C57BL/6J mice or human sWAT wherever mentioned, and IRX3 shRNA were introduced to cells 24 h after seeding. Data were presented as mean ± s.e.m. * P

    Article Snippet: Proteins were assessed with the following antibodies: anti-IRX3 antibody (Abcam, ab25703), anti-GFP antibody (Cell Signaling Technology, 2956s), anti-actin antibody (Santa Cruz Biotechnology, sc-8432), anti-UCP1 antibody (Alpha Diagnostic, ucp1-a), anti-HSP90 antibody (Cell Signaling Technology, 4877s), anti-PGC-1α antibody (Millipore, ab3242), anti-AP2 antibody (Cell Signaling Technology, 3544s), and horseradish peroxidase-conjugated (HRP)-linked secondary antibody (Cell Signaling Technology, 7076, 7074).

    Techniques: In Vitro, Infection, shRNA, Staining, Concentration Assay, Expressing, Pyrolysis Gas Chromatography, Marker, Activation Assay, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Mutagenesis, Over Expression, Real-time Polymerase Chain Reaction, Binding Assay, Chromatin Immunoprecipitation, Construct, Reporter Assay, Isolation, Mouse Assay

    The expression of Irx3 was correlated with the beige/brown adipocyte associated genes in vitro. (A–G) In fully differentiated beige/brown adipocytes from preadipocytes isolated from IWAT, EWAT, and BAT of C57BL/6J and 129/Sv mice, Ucp1 mRNA expression was correlated with Pgc-1α (A) and Dio2 (B). Irx3 mRNA expression showed positive correlation with mRNA levels of (C) Ucp1 , (D) Pgc-1α , (E) Dio2 , and (F) Cox7a1 . Irx3 mRNA expression showed no correlation with Leptin (G). Cells from C57BL/6J were shown in red, and from 129/Sv were in blue. Cells from IWAT, EWAT, and BAT were shown as dot, triangle, and square, respectively. The correlation was analyzed using ΔCT. (H-J) (H) Irx3 (I) Ucp1 and (J) Pparγ mRNA expression during the time course of beige adipocyte differentiation cultures ( n = 3). Quantitative amounts of gene expression were normalized to the housekeeping gene 36b4 . (K–L) Irx3 and Ucp1 protein levels at different time points during beige adipocyte differentiation from IWAT SVFs (K), during brown adipocyte differentiation from BAT SVFs (L). (M) Protein expression of Irx3 and Ap2 at different time points during white adipocyte differentiation from IWAT SVFs. (N) Intracellular location of Ucp1 and Irx3 protein in the fully differentiated beige adipocytes. Ucp1 protein, Irx3 protein and nucleus were indicated in green, red, and blue, respectively. Scar bar, 20 μm. Data were presented as mean ± s.e.m. * P

    Journal: EBioMedicine

    Article Title: IRX3 Promotes the Browning of White Adipocytes and Its Rare Variants are Associated with Human Obesity Risk

    doi: 10.1016/j.ebiom.2017.09.010

    Figure Lengend Snippet: The expression of Irx3 was correlated with the beige/brown adipocyte associated genes in vitro. (A–G) In fully differentiated beige/brown adipocytes from preadipocytes isolated from IWAT, EWAT, and BAT of C57BL/6J and 129/Sv mice, Ucp1 mRNA expression was correlated with Pgc-1α (A) and Dio2 (B). Irx3 mRNA expression showed positive correlation with mRNA levels of (C) Ucp1 , (D) Pgc-1α , (E) Dio2 , and (F) Cox7a1 . Irx3 mRNA expression showed no correlation with Leptin (G). Cells from C57BL/6J were shown in red, and from 129/Sv were in blue. Cells from IWAT, EWAT, and BAT were shown as dot, triangle, and square, respectively. The correlation was analyzed using ΔCT. (H-J) (H) Irx3 (I) Ucp1 and (J) Pparγ mRNA expression during the time course of beige adipocyte differentiation cultures ( n = 3). Quantitative amounts of gene expression were normalized to the housekeeping gene 36b4 . (K–L) Irx3 and Ucp1 protein levels at different time points during beige adipocyte differentiation from IWAT SVFs (K), during brown adipocyte differentiation from BAT SVFs (L). (M) Protein expression of Irx3 and Ap2 at different time points during white adipocyte differentiation from IWAT SVFs. (N) Intracellular location of Ucp1 and Irx3 protein in the fully differentiated beige adipocytes. Ucp1 protein, Irx3 protein and nucleus were indicated in green, red, and blue, respectively. Scar bar, 20 μm. Data were presented as mean ± s.e.m. * P

    Article Snippet: Proteins were assessed with the following antibodies: anti-IRX3 antibody (Abcam, ab25703), anti-GFP antibody (Cell Signaling Technology, 2956s), anti-actin antibody (Santa Cruz Biotechnology, sc-8432), anti-UCP1 antibody (Alpha Diagnostic, ucp1-a), anti-HSP90 antibody (Cell Signaling Technology, 4877s), anti-PGC-1α antibody (Millipore, ab3242), anti-AP2 antibody (Cell Signaling Technology, 3544s), and horseradish peroxidase-conjugated (HRP)-linked secondary antibody (Cell Signaling Technology, 7076, 7074).

    Techniques: Expressing, In Vitro, Isolation, Mouse Assay, Pyrolysis Gas Chromatography

    Impaired diet-induced but not cold exposure-induced brown adipose tissue thermogenesis in Npffr2 − /− mice. a , b Skin temperature at lumbar back ( T Back ) and brown adipose tissue ( T BAT ) by infrared imaging in chow-fed WT ( n = 7) and Npffr2 − /− ( n = 6) mice at 14 weeks of age housed at room temperature (RT 22 °C). b Temperature difference (Δ T BAT-Back ) between T BAT and T Back in chow-fed WT ( n = 7) and Npffr2 − /− ( n = 6) mice. c , d Effect of 6-weeks HFD on T BAT , T Back and Δ T BAT-Back in WT (chow n = 7, HFD n = 6) and Npffr2 − /− (chow n = 4, HFD n = 6) mice. e , f BAT weight and protein content in mice on chow (WT n = 8, Npffr2 − /− n = 8) or 6 weeks of HFD (WT n = 8, Npffr2 − /− n = 11) at 20 weeks of age. g , h BAT UCP-1 and PGC-1α protein expression per mg protein. N values as in e , f . i Representative western blotting image of g , h . j , k BAT UCP-1 and PGC-1α protein expression per BAT depot. N values as in e , f . l , m Effect of 3 weeks single housing (SH) as mild cold exposure vs. group housing (GH) on T BAT , T Back and Δ T BAT-Back in WT (GH n = 7, SH n = 8) and Npffr2 − /− (GH n = 5, SH n = 7) mice. n Effect of RT (22 °C) as mild cold exposure vs. thermoneutrality (28 °C, TN) on T BAT , T Back and Δ T BAT-Back in WT (RT n = 7, TN n = 4) and Npffr2 − /− (RT n = 4, TN n = 4) mice. o Body weight of WT ( n = 7) and Npffr2 − /− ( n = 5) mice fed on HFD for 7 weeks at TN. p , q Weights of white adipose tissue depots of WT ( n = 7) and Npffr2 − /− ( n = 5) mice fed on HFD for 7 weeks at TN; i, e, m and r stand for inguinal, epididymal, mesenteric and retroperitoneal white adipose tissue, respectively. r , s Effect of 7 weeks of HFD on T BAT , T Back and Δ T BAT-Back in WT (Chow n = 4, HFD n = 7) and Npffr2 − /− (Chow n = 4, HFD n = 5) mice at TN. Data are mean ± SEM. One-way ANOVA or repeated measures (o) were used to determine genotype, diet or treatment effects among groups. ∗∗ p

    Journal: Nature Communications

    Article Title: Diet-induced adaptive thermogenesis requires neuropeptide FF receptor-2 signalling

    doi: 10.1038/s41467-018-06462-0

    Figure Lengend Snippet: Impaired diet-induced but not cold exposure-induced brown adipose tissue thermogenesis in Npffr2 − /− mice. a , b Skin temperature at lumbar back ( T Back ) and brown adipose tissue ( T BAT ) by infrared imaging in chow-fed WT ( n = 7) and Npffr2 − /− ( n = 6) mice at 14 weeks of age housed at room temperature (RT 22 °C). b Temperature difference (Δ T BAT-Back ) between T BAT and T Back in chow-fed WT ( n = 7) and Npffr2 − /− ( n = 6) mice. c , d Effect of 6-weeks HFD on T BAT , T Back and Δ T BAT-Back in WT (chow n = 7, HFD n = 6) and Npffr2 − /− (chow n = 4, HFD n = 6) mice. e , f BAT weight and protein content in mice on chow (WT n = 8, Npffr2 − /− n = 8) or 6 weeks of HFD (WT n = 8, Npffr2 − /− n = 11) at 20 weeks of age. g , h BAT UCP-1 and PGC-1α protein expression per mg protein. N values as in e , f . i Representative western blotting image of g , h . j , k BAT UCP-1 and PGC-1α protein expression per BAT depot. N values as in e , f . l , m Effect of 3 weeks single housing (SH) as mild cold exposure vs. group housing (GH) on T BAT , T Back and Δ T BAT-Back in WT (GH n = 7, SH n = 8) and Npffr2 − /− (GH n = 5, SH n = 7) mice. n Effect of RT (22 °C) as mild cold exposure vs. thermoneutrality (28 °C, TN) on T BAT , T Back and Δ T BAT-Back in WT (RT n = 7, TN n = 4) and Npffr2 − /− (RT n = 4, TN n = 4) mice. o Body weight of WT ( n = 7) and Npffr2 − /− ( n = 5) mice fed on HFD for 7 weeks at TN. p , q Weights of white adipose tissue depots of WT ( n = 7) and Npffr2 − /− ( n = 5) mice fed on HFD for 7 weeks at TN; i, e, m and r stand for inguinal, epididymal, mesenteric and retroperitoneal white adipose tissue, respectively. r , s Effect of 7 weeks of HFD on T BAT , T Back and Δ T BAT-Back in WT (Chow n = 4, HFD n = 7) and Npffr2 − /− (Chow n = 4, HFD n = 5) mice at TN. Data are mean ± SEM. One-way ANOVA or repeated measures (o) were used to determine genotype, diet or treatment effects among groups. ∗∗ p

    Article Snippet: Equal amounts of tissue lysate (20 µg protein) were resolved by SDS-polyacrylamide gel electrophoresis and immunoblotted with appropriate antibodies against PGC-1α (Millipore AB3242, 1:1000), UCP-1 (Alpha Diagnostics International, UCP11-A, 1:1000) and GAPDH (Cell Signalling #3683, 1:1000).

    Techniques: Mouse Assay, Imaging, Pyrolysis Gas Chromatography, Expressing, Western Blot