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  • 95
    Millipore anti pgc 1α
    TWEAK inhibits the expression of <t>PGC-1α</t> in cultured myotubes. A ) C2C12 myotubes were treated with indicated amounts of soluble TWEAK protein for 24 h, and mRNA levels of PGC-1α, PGC-1β, Cox I, Cox IV, Cyt c, Tfb2m, Tfam, ATP5b,
    Anti Pgc 1α, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 221 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam pgc 1α
    TGR5 is required for cold-induced scWAT beiging. a Body weight gain of TGR5 wild-type ( Tgr5 +/+ ) and germline TGR5 knockout ( Tgr5 −/− ) mice housed at thermoneutrality (30 °C) or exposed to cold (8 °C) for 7 days. n = 10 per group. b scWAT over body weight (BW) ratio of mice described in a . c – e mRNA levels of beige remodelling markers Ucp1 ( c ), Cidea ( d ), Pgc1a , Tbx1 , Prdm16 , Cd137 , Pparg2 and Cebpb ( e ) in the scWAT of mice described in a . f Representative ( n = 10 per group) western blot of <t>PGC-1α,</t> the mitochondrial marker VDAC1 and beiging markers TBX1 and UCP1 from the scWAT of cold-exposed mice described in a . GAPDH was used as loading control. g Quantitative densitometry of the western blots showed in f . h Representative ( n = 10 per group) western blot of mitochondrial OXPHOS complexes (CII–CV) from the scWAT of cold-exposed mice described in a , GAPDH was used as loading control. i Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the scWAT of cold-exposed mice described in a . j , k Representative haematoxylin and eosin ( j ) and UCP1 ( k ) stainings of scWAT sections from cold-exposed mice described in a . Scale bars = 50 μm. Results represent mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01 and *** P ≤ 0.001 vs. Tgr5 +/+ group (at 30 and/or 8 °C) by one-way ANOVA followed by Bonferroni post hoc test ( a – e ) or Student’s t -test ( g , i A–C
    Pgc 1α, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1241 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Cell Signaling Technology Inc pgc 1α
    Knockdown of PRL-3 decreases the expression of <t>PGC-1α</t> targeted SOD2 and UCP2. Notes: ( A ) Western blot (up) and RT-PCR (bottom) results showed that knockdown of RAP1 decreased the expression of PGC-1α. ( B ) Western blot (up) and RT-PCR (bottom) results showed that knockdown of PRL-3 decreased the expression of PGC-1α. ( C ) Knockdown of RAP1 in PRL-3 overexpressing cells suppressed the upregulation of PGC-1α. PRL-3 was overexpressed through transfection of pcDNA3-PRL-3-myc, and pcDNA3-vector-myc as control. Then RAP1 was knockdown through shRNA lentivirus infection. ( D ) Measurement of relative mtDNA by RT-PCR after knockdown of either PRL-3 or RAP1. ( E ) Knockdown of either RAP1 (up) or PRL-3 (bottom) decreased the mRNA level of SOD2 and UCP2. ( F ) Gene expression correlation analysis between PRL-3 and SOD2, RAP1 and SOD2 in TCGA data (PRL-3-SOD2: P =0.0012, R =0.49; RAP1-SOD2: P
    Pgc 1α, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 443 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Millipore pgc 1α
    A Schematic depiction of the mechanism of CD105/BMP signaling in conferring radiation resistance. Radiation of prostate cancer results in upregulation of cell surface CD105 expression. The ensuing CD105/BMP signaling is essential and necessary for SIRT1 expression and downstream activity. SIRT1 facilitates DNA damage repair, destabilizes p53, and activates of <t>PGC-1α</t> in response to radiation. Consequently, CD105 can regulate glycolysis and mitochondrial biogenesis to meet the higher energy demands necessary for survival following irradiation. The loss of functional p53 enables glycolysis as a source of ATP generation and survival
    Pgc 1α, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 900 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Novus Biologicals pgc1 alpha antibody
    A Schematic depiction of the mechanism of CD105/BMP signaling in conferring radiation resistance. Radiation of prostate cancer results in upregulation of cell surface CD105 expression. The ensuing CD105/BMP signaling is essential and necessary for SIRT1 expression and downstream activity. SIRT1 facilitates DNA damage repair, destabilizes p53, and activates of <t>PGC-1α</t> in response to radiation. Consequently, CD105 can regulate glycolysis and mitochondrial biogenesis to meet the higher energy demands necessary for survival following irradiation. The loss of functional p53 enables glycolysis as a source of ATP generation and survival
    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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    99
    Millipore anti pgc 1α antibody
    A Schematic depiction of the mechanism of CD105/BMP signaling in conferring radiation resistance. Radiation of prostate cancer results in upregulation of cell surface CD105 expression. The ensuing CD105/BMP signaling is essential and necessary for SIRT1 expression and downstream activity. SIRT1 facilitates DNA damage repair, destabilizes p53, and activates of <t>PGC-1α</t> in response to radiation. Consequently, CD105 can regulate glycolysis and mitochondrial biogenesis to meet the higher energy demands necessary for survival following irradiation. The loss of functional p53 enables glycolysis as a source of ATP generation and survival
    Anti Pgc 1α Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 115 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc pgc1α
    The effect of BBR addition in the preservation solution before ischemia on the gene expression of Sirt 1 and 3, <t>PGC-1α,</t> MnSOD2, COX IV and NDUFS8 as evaluated by qPCR. Tissue samples were obtained from animals sacrificed after 12 h of hepatic exposure to preservation solution and 1 h of reperfusion, as described in the Materials and Methods Section. Control animals were sham operated. I/R livers (Group B) had a reduced gene expression of SirT1, the nuclearly-encoded mitochondrial respiratory complex IV subunit IV (COX IV) and PGC-1α, which were prevented by BBR pre-treatment (Group A). No differences were found in the gene expression of SirT3, the mitochondrial antioxidant protein MnSOD2 and the mitochondrially-encoded mitochondrial respiratory complex I iron-sulfur protein 8 (NDUFS8). Boxes represent the means ± 5–95 percentile of an n = 6 different experiments. * indicates a statistically-significant difference ( p
    Pgc1α, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 193 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc anti pgc1α
    METRNL alleviates inflammation and insulin resistance through a PPARδ-mediated pathway. a Western blot analysis of PPARδ expression in differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. b Confirmation of PPARδ siRNA efficiency in differentiated C2C12 cells. Western blot analysis of palmitate (200 μM)-induced inflammation markers in PPARα siRNA (20 nM)-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. c Western blot analysis of the palmitate-induced impairment of IRS-1 and Akt phosphorylation in PPARδ siRNA-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. Human insulin (10 nM) was used to stimulate insulin signaling for 3 min. d Western blot analysis of PPARδ expression in soleus muscle of mice treated with HFD and METRNL (5 animals/treatment group). e Western blot analysis of METRNL (200 ng/mL)-induced <t>PGC1α</t> expression in PPARδ siRNA (20 nM)-transfected differentiated C2C12 cells for 24 h. Western blot analysis of f AMPK phosphorylation in PPARδ siRNA or g PPARδ expression in AMPK siRNA-transfected differentiated C2C12 cells treated with METRNL (200 ng/mL) for 24 h. Confirmation of PPARδ siRNA efficiency in skeletal muscle of mice. Western blot analysis of IRS-1 and Akt phosphorylation ( h ), and inflammatory markers ( i ) in PPARδ siRNA-transfected skeletal muscle of experimental mice. j Western blot analysis of PPARδ expression in AMPK siRNA-transfected skeletal muscle of experimental mice. k Western blot analysis of AMPK phosphorylation in PPARδ siRNA-transfected skeletal muscle of experimental mice. Means ± SEM were obtained from three separate experiments or five animals. *** P
    Anti Pgc1α, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Abcam rabbit anti pgc 1α
    <t>PGC-1α</t> knockdown inhibits T3-induced dendritic growth in Purkinje cells. (A) HEK293T cells expressing mouse PGC-1α were transfected with PGC-1α-scramble (scr) or short hairpin RNA (shRNA) construct and analyzed by western blotting with anti-PGC-1α and anti-β-actin antibodies. (B,C) Cultured Purkinje cells were transfected with PGC-1α-shRNA or scr-shRNA (control) constructs and stained for Calbindin and PGC-1α (B) or Calbindin and cytochrome C oxidase IV (COX-IV) (C) at 10 DIV. Scale bars, 10 μm. (D) Representative images of Purkinje cells transfected with scr shRNA (control), PGC-1α shRNA (PGC-1α shRNA), or PGC-1α shRNA plus an shRNA-resistant mutant of PGC-1α (shRNA + rescue). Cells were cultured in the presence of 10 nM T3 until 10 DIV and immunostained with anti-Calbindin and anti-PDH antibodies. Boxed regions in the upper panels are enlarged in lower panels. Scale bars, 20 μm. (E–G) Quantitative analyses of the total dendritic length (E) , number of dendritic branches (F) and PDH signal (G) . Data represent mean ± SEM, N = 30 for each point, ** p
    Rabbit Anti Pgc 1α, supplied by Abcam, used in various techniques. Bioz Stars score: 88/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Millipore mouse anti pgc 1α
    <t>PGC-1α</t> knockdown inhibits T3-induced dendritic growth in Purkinje cells. (A) HEK293T cells expressing mouse PGC-1α were transfected with PGC-1α-scramble (scr) or short hairpin RNA (shRNA) construct and analyzed by western blotting with anti-PGC-1α and anti-β-actin antibodies. (B,C) Cultured Purkinje cells were transfected with PGC-1α-shRNA or scr-shRNA (control) constructs and stained for Calbindin and PGC-1α (B) or Calbindin and cytochrome C oxidase IV (COX-IV) (C) at 10 DIV. Scale bars, 10 μm. (D) Representative images of Purkinje cells transfected with scr shRNA (control), PGC-1α shRNA (PGC-1α shRNA), or PGC-1α shRNA plus an shRNA-resistant mutant of PGC-1α (shRNA + rescue). Cells were cultured in the presence of 10 nM T3 until 10 DIV and immunostained with anti-Calbindin and anti-PDH antibodies. Boxed regions in the upper panels are enlarged in lower panels. Scale bars, 20 μm. (E–G) Quantitative analyses of the total dendritic length (E) , number of dendritic branches (F) and PDH signal (G) . Data represent mean ± SEM, N = 30 for each point, ** p
    Mouse Anti Pgc 1α, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Abcam pgc1alpha
    Alpha-synuclein binds to <t>PGC1alpha</t> promoter sequences in vitro and in vivo and in PD brain tissues
    Pgc1alpha, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Santa Cruz Biotechnology pgc 1α antibody
    ( A ) Representative Western blot of p-p65 (Ser536), p65, p-STAT3 (Tyr705) and STAT3 in the livers of the sham <t>PGC-1α</t> +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). GAPDH was used as the loading control. ( B ) Representative Western blot of p65 and PGC-1α in the PGC-1α immunoprecipitate of the livers of the sham PGC-1α +/+ (WT) mice and at 1 h after cerulein-induced AP mice. ( C ) Representative Western blot of p-STAT3 (Tyr705) and p65 in the p65 immunoprecipitate of the livers of the PGC-1α +/+ (WT) mice and PGC-1α -/- (KO) mice with pancreatitis (Cerulein). IgG was used as the loading control. There were six mice per group.
    Pgc 1α Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 88 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Biogenesis Inc pgc 1α
    Proposed model whereby AngII infusion leads to mitochondrial dysfunction and skeletal muscle wasting. Parallel to the well-characterized AngII-mediated activation of the FOXO-E3-UPS axis via inhibition of Akt, AngII also induces expression of the phosphatase PP2Cα, which dephosphorylates and inactivates AMPK. This leads to reduced <t>PGC-1α,</t> NRF1, and TFAM expression (less mitochondrial biogenesis), and reduced ULK1 activity. The AngII-mediated reduction in ULK1 activation inhibits a critical early step in the autophagy pathway and prevents recycling of damaged mitochondria (mitophagy). AngII also inhibits both mitochondrial fission and fusion through predominately AMPK-independent pathways, which likely contribute to mitochondrial dysfunction caused by elevated AngII. Prolonged mitochondrial dysfunction and energy depletion ultimately leads to release of caspase-3, initiation of apoptosis, and wasting.
    Pgc 1α, supplied by Biogenesis Inc, used in various techniques. Bioz Stars score: 92/100, based on 63 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    TWEAK inhibits the expression of PGC-1α in cultured myotubes. A ) C2C12 myotubes were treated with indicated amounts of soluble TWEAK protein for 24 h, and mRNA levels of PGC-1α, PGC-1β, Cox I, Cox IV, Cyt c, Tfb2m, Tfam, ATP5b,

    Journal: The FASEB Journal

    Article Title: Regulatory circuitry of TWEAK-Fn14 system and PGC-1α in skeletal muscle atrophy program

    doi: 10.1096/fj.13-242123

    Figure Lengend Snippet: TWEAK inhibits the expression of PGC-1α in cultured myotubes. A ) C2C12 myotubes were treated with indicated amounts of soluble TWEAK protein for 24 h, and mRNA levels of PGC-1α, PGC-1β, Cox I, Cox IV, Cyt c, Tfb2m, Tfam, ATP5b,

    Article Snippet: Approximately 100 μg protein was resolved on each lane on 10 or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electrotransferred onto nitrocellulose or polyvinylidene difluoride membrane, and probed using anti-PGC-1α (1:500, cat. no. ST1203; Calbiochem, La Jolla, CA, USA), Anti-total oxphos cocktail (1:500, ab110413; Abcam, Cambridge, MA, USA), anti-phospho-p65 (1:1000, 3033; Cell Signaling Technology, Danvers, MA, USA), anti-p65 (1:1000, 8242; Cell Signaling Technology), anti-p100/p52 (1:1000, 4882; Cell Signaling Technology), and anti-Fn14 (1:500, 4403; Cell Signaling Technology) and detected by enhanced chemiluminescence.

    Techniques: Expressing, Pyrolysis Gas Chromatography, Cell Culture

    Tg overexpression of PGC-1α inhibits the progressive muscle atrophy in TWEAK-Tg mice. A ) TA muscle isolated from 15-mo-old littermate WT, TWEAK-Tg, PGC-1α-Tg, and TWEAK-PGC-1α double-Tg mice were processed for H E staining.

    Journal: The FASEB Journal

    Article Title: Regulatory circuitry of TWEAK-Fn14 system and PGC-1α in skeletal muscle atrophy program

    doi: 10.1096/fj.13-242123

    Figure Lengend Snippet: Tg overexpression of PGC-1α inhibits the progressive muscle atrophy in TWEAK-Tg mice. A ) TA muscle isolated from 15-mo-old littermate WT, TWEAK-Tg, PGC-1α-Tg, and TWEAK-PGC-1α double-Tg mice were processed for H E staining.

    Article Snippet: Approximately 100 μg protein was resolved on each lane on 10 or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electrotransferred onto nitrocellulose or polyvinylidene difluoride membrane, and probed using anti-PGC-1α (1:500, cat. no. ST1203; Calbiochem, La Jolla, CA, USA), Anti-total oxphos cocktail (1:500, ab110413; Abcam, Cambridge, MA, USA), anti-phospho-p65 (1:1000, 3033; Cell Signaling Technology, Danvers, MA, USA), anti-p65 (1:1000, 8242; Cell Signaling Technology), anti-p100/p52 (1:1000, 4882; Cell Signaling Technology), and anti-Fn14 (1:500, 4403; Cell Signaling Technology) and detected by enhanced chemiluminescence.

    Techniques: Over Expression, Pyrolysis Gas Chromatography, Mouse Assay, Isolation, Staining

    Overexpression of PGC-1α inhibits TWEAK-induced atrophy in cultured muscle cells. Myofiber cultures established from EDL muscle of WT and PGC-1α Tg mice were treated with indicated concentration of soluble TWEAK protein for 72 h. A ) Representative

    Journal: The FASEB Journal

    Article Title: Regulatory circuitry of TWEAK-Fn14 system and PGC-1α in skeletal muscle atrophy program

    doi: 10.1096/fj.13-242123

    Figure Lengend Snippet: Overexpression of PGC-1α inhibits TWEAK-induced atrophy in cultured muscle cells. Myofiber cultures established from EDL muscle of WT and PGC-1α Tg mice were treated with indicated concentration of soluble TWEAK protein for 72 h. A ) Representative

    Article Snippet: Approximately 100 μg protein was resolved on each lane on 10 or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electrotransferred onto nitrocellulose or polyvinylidene difluoride membrane, and probed using anti-PGC-1α (1:500, cat. no. ST1203; Calbiochem, La Jolla, CA, USA), Anti-total oxphos cocktail (1:500, ab110413; Abcam, Cambridge, MA, USA), anti-phospho-p65 (1:1000, 3033; Cell Signaling Technology, Danvers, MA, USA), anti-p65 (1:1000, 8242; Cell Signaling Technology), anti-p100/p52 (1:1000, 4882; Cell Signaling Technology), and anti-Fn14 (1:500, 4403; Cell Signaling Technology) and detected by enhanced chemiluminescence.

    Techniques: Over Expression, Pyrolysis Gas Chromatography, Cell Culture, Mouse Assay, Concentration Assay

    TWEAK represses PGC-1α levels in denervated skeletal muscle of mice. Quantitative estimation of mRNA levels of different genes in TA muscle by QRT-PCR assay. A ) Relative mRNA levels of TWEAK receptor Fn14 and PGC-1α in undenervated and

    Journal: The FASEB Journal

    Article Title: Regulatory circuitry of TWEAK-Fn14 system and PGC-1α in skeletal muscle atrophy program

    doi: 10.1096/fj.13-242123

    Figure Lengend Snippet: TWEAK represses PGC-1α levels in denervated skeletal muscle of mice. Quantitative estimation of mRNA levels of different genes in TA muscle by QRT-PCR assay. A ) Relative mRNA levels of TWEAK receptor Fn14 and PGC-1α in undenervated and

    Article Snippet: Approximately 100 μg protein was resolved on each lane on 10 or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electrotransferred onto nitrocellulose or polyvinylidene difluoride membrane, and probed using anti-PGC-1α (1:500, cat. no. ST1203; Calbiochem, La Jolla, CA, USA), Anti-total oxphos cocktail (1:500, ab110413; Abcam, Cambridge, MA, USA), anti-phospho-p65 (1:1000, 3033; Cell Signaling Technology, Danvers, MA, USA), anti-p65 (1:1000, 8242; Cell Signaling Technology), anti-p100/p52 (1:1000, 4882; Cell Signaling Technology), and anti-Fn14 (1:500, 4403; Cell Signaling Technology) and detected by enhanced chemiluminescence.

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

    Overexpression of PGC-1α blunts the expression of atrogenes and augments the levels of mitochondria-related genes in TWEAK-treated myotubes. Primary myoblasts prepared from littermate WT and PGC-1α Tg mice were differentiated into myotubes,

    Journal: The FASEB Journal

    Article Title: Regulatory circuitry of TWEAK-Fn14 system and PGC-1α in skeletal muscle atrophy program

    doi: 10.1096/fj.13-242123

    Figure Lengend Snippet: Overexpression of PGC-1α blunts the expression of atrogenes and augments the levels of mitochondria-related genes in TWEAK-treated myotubes. Primary myoblasts prepared from littermate WT and PGC-1α Tg mice were differentiated into myotubes,

    Article Snippet: Approximately 100 μg protein was resolved on each lane on 10 or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electrotransferred onto nitrocellulose or polyvinylidene difluoride membrane, and probed using anti-PGC-1α (1:500, cat. no. ST1203; Calbiochem, La Jolla, CA, USA), Anti-total oxphos cocktail (1:500, ab110413; Abcam, Cambridge, MA, USA), anti-phospho-p65 (1:1000, 3033; Cell Signaling Technology, Danvers, MA, USA), anti-p65 (1:1000, 8242; Cell Signaling Technology), anti-p100/p52 (1:1000, 4882; Cell Signaling Technology), and anti-Fn14 (1:500, 4403; Cell Signaling Technology) and detected by enhanced chemiluminescence.

    Techniques: Over Expression, Pyrolysis Gas Chromatography, Expressing, Mouse Assay

    Overexpression of PGC-1α inhibits the expression of Fn14 in denervated muscle of mice. A ) Relative mRNA levels of Fn14, TWEAK, and MuRF1 in undenervated and denervated TA muscle of WT and PGC-1α-Tg mice, measured by performing QRT-PCR

    Journal: The FASEB Journal

    Article Title: Regulatory circuitry of TWEAK-Fn14 system and PGC-1α in skeletal muscle atrophy program

    doi: 10.1096/fj.13-242123

    Figure Lengend Snippet: Overexpression of PGC-1α inhibits the expression of Fn14 in denervated muscle of mice. A ) Relative mRNA levels of Fn14, TWEAK, and MuRF1 in undenervated and denervated TA muscle of WT and PGC-1α-Tg mice, measured by performing QRT-PCR

    Article Snippet: Approximately 100 μg protein was resolved on each lane on 10 or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electrotransferred onto nitrocellulose or polyvinylidene difluoride membrane, and probed using anti-PGC-1α (1:500, cat. no. ST1203; Calbiochem, La Jolla, CA, USA), Anti-total oxphos cocktail (1:500, ab110413; Abcam, Cambridge, MA, USA), anti-phospho-p65 (1:1000, 3033; Cell Signaling Technology, Danvers, MA, USA), anti-p65 (1:1000, 8242; Cell Signaling Technology), anti-p100/p52 (1:1000, 4882; Cell Signaling Technology), and anti-Fn14 (1:500, 4403; Cell Signaling Technology) and detected by enhanced chemiluminescence.

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

    PGC-1α inhibits the TWEAK-induced activation of NF-κB in myotubes. Primary myotubes prepared from WT and PGC-1α Tg mice were treated with 100 ng/ml TWEAK protein for indicated time period and analyzed for the activation of NF-κB.

    Journal: The FASEB Journal

    Article Title: Regulatory circuitry of TWEAK-Fn14 system and PGC-1α in skeletal muscle atrophy program

    doi: 10.1096/fj.13-242123

    Figure Lengend Snippet: PGC-1α inhibits the TWEAK-induced activation of NF-κB in myotubes. Primary myotubes prepared from WT and PGC-1α Tg mice were treated with 100 ng/ml TWEAK protein for indicated time period and analyzed for the activation of NF-κB.

    Article Snippet: Approximately 100 μg protein was resolved on each lane on 10 or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electrotransferred onto nitrocellulose or polyvinylidene difluoride membrane, and probed using anti-PGC-1α (1:500, cat. no. ST1203; Calbiochem, La Jolla, CA, USA), Anti-total oxphos cocktail (1:500, ab110413; Abcam, Cambridge, MA, USA), anti-phospho-p65 (1:1000, 3033; Cell Signaling Technology, Danvers, MA, USA), anti-p65 (1:1000, 8242; Cell Signaling Technology), anti-p100/p52 (1:1000, 4882; Cell Signaling Technology), and anti-Fn14 (1:500, 4403; Cell Signaling Technology) and detected by enhanced chemiluminescence.

    Techniques: Pyrolysis Gas Chromatography, Activation Assay, Mouse Assay

    TGR5 is required for cold-induced scWAT beiging. a Body weight gain of TGR5 wild-type ( Tgr5 +/+ ) and germline TGR5 knockout ( Tgr5 −/− ) mice housed at thermoneutrality (30 °C) or exposed to cold (8 °C) for 7 days. n = 10 per group. b scWAT over body weight (BW) ratio of mice described in a . c – e mRNA levels of beige remodelling markers Ucp1 ( c ), Cidea ( d ), Pgc1a , Tbx1 , Prdm16 , Cd137 , Pparg2 and Cebpb ( e ) in the scWAT of mice described in a . f Representative ( n = 10 per group) western blot of PGC-1α, the mitochondrial marker VDAC1 and beiging markers TBX1 and UCP1 from the scWAT of cold-exposed mice described in a . GAPDH was used as loading control. g Quantitative densitometry of the western blots showed in f . h Representative ( n = 10 per group) western blot of mitochondrial OXPHOS complexes (CII–CV) from the scWAT of cold-exposed mice described in a , GAPDH was used as loading control. i Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the scWAT of cold-exposed mice described in a . j , k Representative haematoxylin and eosin ( j ) and UCP1 ( k ) stainings of scWAT sections from cold-exposed mice described in a . Scale bars = 50 μm. Results represent mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01 and *** P ≤ 0.001 vs. Tgr5 +/+ group (at 30 and/or 8 °C) by one-way ANOVA followed by Bonferroni post hoc test ( a – e ) or Student’s t -test ( g , i A–C

    Journal: Nature Communications

    Article Title: TGR5 signalling promotes mitochondrial fission and beige remodelling of white adipose tissue

    doi: 10.1038/s41467-017-02068-0

    Figure Lengend Snippet: TGR5 is required for cold-induced scWAT beiging. a Body weight gain of TGR5 wild-type ( Tgr5 +/+ ) and germline TGR5 knockout ( Tgr5 −/− ) mice housed at thermoneutrality (30 °C) or exposed to cold (8 °C) for 7 days. n = 10 per group. b scWAT over body weight (BW) ratio of mice described in a . c – e mRNA levels of beige remodelling markers Ucp1 ( c ), Cidea ( d ), Pgc1a , Tbx1 , Prdm16 , Cd137 , Pparg2 and Cebpb ( e ) in the scWAT of mice described in a . f Representative ( n = 10 per group) western blot of PGC-1α, the mitochondrial marker VDAC1 and beiging markers TBX1 and UCP1 from the scWAT of cold-exposed mice described in a . GAPDH was used as loading control. g Quantitative densitometry of the western blots showed in f . h Representative ( n = 10 per group) western blot of mitochondrial OXPHOS complexes (CII–CV) from the scWAT of cold-exposed mice described in a , GAPDH was used as loading control. i Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the scWAT of cold-exposed mice described in a . j , k Representative haematoxylin and eosin ( j ) and UCP1 ( k ) stainings of scWAT sections from cold-exposed mice described in a . Scale bars = 50 μm. Results represent mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01 and *** P ≤ 0.001 vs. Tgr5 +/+ group (at 30 and/or 8 °C) by one-way ANOVA followed by Bonferroni post hoc test ( a – e ) or Student’s t -test ( g , i A–C

    Article Snippet: PGC-1α (ab54481 1:1000), UCP1 (ab10983 1:1000 for animal tissues, 1:500 for primary culture), TBX1 (ab109313 1:1000) and VDAC1 (ab14734 1:1000) antibodies were from Abcam; anti-CREB (9197 1:1000), p-CREB Ser133 (9191 1:1000), ERK (4695 1:1000), p-ERK (4376 1:1000), DRP1 (4E11B11 1:1000), p-DRP1 Ser616 (3455 1:1000) and p-DRP1 Ser637 (4867S 1:1000) antibodies were from Cell Signalling; anti-MFF antibody (17090-1-AP 1:1000) was from Proteintech group; anti-PARP1 (sc-7150 1:1000), GAPDH (sc-47724 1:2000) and TOMM40 (sc-11414 1:1000) antibodies were from Santa Cruz Biotechnology.

    Techniques: Knock-Out, Mouse Assay, Western Blot, Pyrolysis Gas Chromatography, Marker

    Adipocyte TGR5 is required for cold-induced scWAT beiging. a Body temperature of control mice ( Tgr5 Adipoq+/+ ) and WAT-specific TGR5 knockout ( Tgr5 Adipoq−/− ) mice exposed to cold (8 °C) for 7 days. n = 10 per group. b Representative ( n = 5 per group) haematoxylin and eosin stainings of scWAT of mice described in a . c – e mRNA levels of beige remodelling markers Ucp1 ( c ) Pgc1a , Tbx1 , Cidea and Pparg2 ( d ), and Prdm16 , Cd137 and Cebpb ( e ) in the scWAT of Tgr5 Adipoq+/+ and Tgr5 Adipoq−/− mice housed at thermoneutrality (30 °C) or exposed to cold (8 °C) for 7 days. n = 10 per group. f Representative ( n = 10 per group) western blot of PGC-1α, the mitochondrial marker VDAC1 and beiging markers TBX1 and UCP1 from the scWAT of mice described in a . GAPDH was used as loading control. g Quantitative densitometry of the western blots showed in f . h Representative ( n = 5 per group) UCP1 staining of scWAT sections from mice described in a . Scale bars = 50 μm. i Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the scWAT of mice described in a . Results represent mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01 and *** P ≤ 0.001 vs. Tgr5 Adipoq+/+ group (at 30 and/or 8 °C) by one-way ANOVA ( c – e ) and two-way ANOVA ( a ) followed by Bonferroni post hoc test or Student’s t test ( g , i A–E

    Journal: Nature Communications

    Article Title: TGR5 signalling promotes mitochondrial fission and beige remodelling of white adipose tissue

    doi: 10.1038/s41467-017-02068-0

    Figure Lengend Snippet: Adipocyte TGR5 is required for cold-induced scWAT beiging. a Body temperature of control mice ( Tgr5 Adipoq+/+ ) and WAT-specific TGR5 knockout ( Tgr5 Adipoq−/− ) mice exposed to cold (8 °C) for 7 days. n = 10 per group. b Representative ( n = 5 per group) haematoxylin and eosin stainings of scWAT of mice described in a . c – e mRNA levels of beige remodelling markers Ucp1 ( c ) Pgc1a , Tbx1 , Cidea and Pparg2 ( d ), and Prdm16 , Cd137 and Cebpb ( e ) in the scWAT of Tgr5 Adipoq+/+ and Tgr5 Adipoq−/− mice housed at thermoneutrality (30 °C) or exposed to cold (8 °C) for 7 days. n = 10 per group. f Representative ( n = 10 per group) western blot of PGC-1α, the mitochondrial marker VDAC1 and beiging markers TBX1 and UCP1 from the scWAT of mice described in a . GAPDH was used as loading control. g Quantitative densitometry of the western blots showed in f . h Representative ( n = 5 per group) UCP1 staining of scWAT sections from mice described in a . Scale bars = 50 μm. i Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the scWAT of mice described in a . Results represent mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01 and *** P ≤ 0.001 vs. Tgr5 Adipoq+/+ group (at 30 and/or 8 °C) by one-way ANOVA ( c – e ) and two-way ANOVA ( a ) followed by Bonferroni post hoc test or Student’s t test ( g , i A–E

    Article Snippet: PGC-1α (ab54481 1:1000), UCP1 (ab10983 1:1000 for animal tissues, 1:500 for primary culture), TBX1 (ab109313 1:1000) and VDAC1 (ab14734 1:1000) antibodies were from Abcam; anti-CREB (9197 1:1000), p-CREB Ser133 (9191 1:1000), ERK (4695 1:1000), p-ERK (4376 1:1000), DRP1 (4E11B11 1:1000), p-DRP1 Ser616 (3455 1:1000) and p-DRP1 Ser637 (4867S 1:1000) antibodies were from Cell Signalling; anti-MFF antibody (17090-1-AP 1:1000) was from Proteintech group; anti-PARP1 (sc-7150 1:1000), GAPDH (sc-47724 1:2000) and TOMM40 (sc-11414 1:1000) antibodies were from Santa Cruz Biotechnology.

    Techniques: Mouse Assay, Knock-Out, Western Blot, Pyrolysis Gas Chromatography, Marker, Staining

    TGR5 activation induces scWAT beiging in mice fed a high-fat diet. a Body weight curves of TGR5 wild-type ( Tgr5 +/+ ) fed a high-fat (HF) diet for 20 weeks in the presence or absence of the selective TGR5 agonist INT-777. n = 10 per group. b mRNA levels of beige remodelling markers Pgc1a , Ucp1 , Tbx1 , Prdm16 , Cidea, Cd137 , Pparg2 and Cebpb in the scWAT of mice described in a . c Representative ( n = 10 per group) western blot of PGC-1α, the mitochondrial marker VDAC1, and beiging markers TBX1 and UCP1 from the scWAT of mice described in a . GAPDH was used as loading control. d Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the scWAT of mice described in a . e Representative haematoxylin and eosin staining from the scWAT of mice described in a . f Adipocyte area quantification from images shown in e . g Representative ( n = 5 per group) UCP1 immunostaining of scWAT sections from mice described in a . h Quantification of UCP1 immunostaining intensity depicted in g . Scale bars = 50 μm. Results represent mean ± SEM. * P ≤ 0.05 and ** P ≤ 0.01 vs. Tgr5 +/+ HF group by two-way ANOVA followed by Bonferroni post hoc test ( a ) or Student’s t test ( b , d , h A and B

    Journal: Nature Communications

    Article Title: TGR5 signalling promotes mitochondrial fission and beige remodelling of white adipose tissue

    doi: 10.1038/s41467-017-02068-0

    Figure Lengend Snippet: TGR5 activation induces scWAT beiging in mice fed a high-fat diet. a Body weight curves of TGR5 wild-type ( Tgr5 +/+ ) fed a high-fat (HF) diet for 20 weeks in the presence or absence of the selective TGR5 agonist INT-777. n = 10 per group. b mRNA levels of beige remodelling markers Pgc1a , Ucp1 , Tbx1 , Prdm16 , Cidea, Cd137 , Pparg2 and Cebpb in the scWAT of mice described in a . c Representative ( n = 10 per group) western blot of PGC-1α, the mitochondrial marker VDAC1, and beiging markers TBX1 and UCP1 from the scWAT of mice described in a . GAPDH was used as loading control. d Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the scWAT of mice described in a . e Representative haematoxylin and eosin staining from the scWAT of mice described in a . f Adipocyte area quantification from images shown in e . g Representative ( n = 5 per group) UCP1 immunostaining of scWAT sections from mice described in a . h Quantification of UCP1 immunostaining intensity depicted in g . Scale bars = 50 μm. Results represent mean ± SEM. * P ≤ 0.05 and ** P ≤ 0.01 vs. Tgr5 +/+ HF group by two-way ANOVA followed by Bonferroni post hoc test ( a ) or Student’s t test ( b , d , h A and B

    Article Snippet: PGC-1α (ab54481 1:1000), UCP1 (ab10983 1:1000 for animal tissues, 1:500 for primary culture), TBX1 (ab109313 1:1000) and VDAC1 (ab14734 1:1000) antibodies were from Abcam; anti-CREB (9197 1:1000), p-CREB Ser133 (9191 1:1000), ERK (4695 1:1000), p-ERK (4376 1:1000), DRP1 (4E11B11 1:1000), p-DRP1 Ser616 (3455 1:1000) and p-DRP1 Ser637 (4867S 1:1000) antibodies were from Cell Signalling; anti-MFF antibody (17090-1-AP 1:1000) was from Proteintech group; anti-PARP1 (sc-7150 1:1000), GAPDH (sc-47724 1:2000) and TOMM40 (sc-11414 1:1000) antibodies were from Santa Cruz Biotechnology.

    Techniques: Activation Assay, Mouse Assay, Western Blot, Pyrolysis Gas Chromatography, Marker, Staining, Immunostaining

    TGR5 activation induces scWAT beiging at thermoneutrality. a Body weight gain of TGR5 wild-type ( Tgr5 +/+ ) and germline TGR5 knockout ( Tgr5 −/− ) mice housed at thermoneutrality (30 °C) and subjected to a daily administration of the selective TGR5 agonist INT-777 or vehicle for 7 days. n = 10 per group. b scWAT over body weight (BW) ratio of the mice described in a . c , d mRNA levels of beige remodelling markers Ucp1 and Cidea ( c ), Pgc1a , Tbx1 , Prdm16 , and Cd137 , Pparg2 and Cebpb ; Tgr5 and adrenergic related genes ( Adrb3 and Th ) ( d ) in the scWAT of mice described in a . e Representative ( n = 10 per group) western blot of PGC-1α, the mitochondrial marker VDAC1, and beiging markers TBX1 and UCP1 from the scWAT of mice described in a . GAPDH was used as loading control. f Representative ( n = 5 per group) UCP1 immunostaining of scWAT sections from mice described in a . Scale bars = 50 μm. Results represent mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01 and *** P ≤ 0.001 vs. Tgr5 +/+ A–D

    Journal: Nature Communications

    Article Title: TGR5 signalling promotes mitochondrial fission and beige remodelling of white adipose tissue

    doi: 10.1038/s41467-017-02068-0

    Figure Lengend Snippet: TGR5 activation induces scWAT beiging at thermoneutrality. a Body weight gain of TGR5 wild-type ( Tgr5 +/+ ) and germline TGR5 knockout ( Tgr5 −/− ) mice housed at thermoneutrality (30 °C) and subjected to a daily administration of the selective TGR5 agonist INT-777 or vehicle for 7 days. n = 10 per group. b scWAT over body weight (BW) ratio of the mice described in a . c , d mRNA levels of beige remodelling markers Ucp1 and Cidea ( c ), Pgc1a , Tbx1 , Prdm16 , and Cd137 , Pparg2 and Cebpb ; Tgr5 and adrenergic related genes ( Adrb3 and Th ) ( d ) in the scWAT of mice described in a . e Representative ( n = 10 per group) western blot of PGC-1α, the mitochondrial marker VDAC1, and beiging markers TBX1 and UCP1 from the scWAT of mice described in a . GAPDH was used as loading control. f Representative ( n = 5 per group) UCP1 immunostaining of scWAT sections from mice described in a . Scale bars = 50 μm. Results represent mean ± SEM. * P ≤ 0.05, ** P ≤ 0.01 and *** P ≤ 0.001 vs. Tgr5 +/+ A–D

    Article Snippet: PGC-1α (ab54481 1:1000), UCP1 (ab10983 1:1000 for animal tissues, 1:500 for primary culture), TBX1 (ab109313 1:1000) and VDAC1 (ab14734 1:1000) antibodies were from Abcam; anti-CREB (9197 1:1000), p-CREB Ser133 (9191 1:1000), ERK (4695 1:1000), p-ERK (4376 1:1000), DRP1 (4E11B11 1:1000), p-DRP1 Ser616 (3455 1:1000) and p-DRP1 Ser637 (4867S 1:1000) antibodies were from Cell Signalling; anti-MFF antibody (17090-1-AP 1:1000) was from Proteintech group; anti-PARP1 (sc-7150 1:1000), GAPDH (sc-47724 1:2000) and TOMM40 (sc-11414 1:1000) antibodies were from Santa Cruz Biotechnology.

    Techniques: Activation Assay, Knock-Out, Mouse Assay, Western Blot, Pyrolysis Gas Chromatography, Marker, Immunostaining

    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

    Effect of Honokiol on SIRT3 activation and AMPK-CREB-PGC1α pathway. Effect of increasing concentrations of insulin (0, 1, 5 and 10nM) on TsAβ (Total secreted Aβ) along with the densitometric analysis of band intensity normalized to β-Actin. (B) Representative Western blot of SIRT3 performed on whole cell lysates from PS70 cells exposed to either vehicle (DMSO) or 5 and 10μM Honokiol for 24 and 48 h. The graph displays the densitometric analysis of band intensity of the SIRT3 normalized to the corresponding GADPH level, used as loading control. (C) Effect of Insulin 10nM and Honokiol (5 and 10μM) on p-AMPK/AMPK ratio. Representative Western blot of total AMPK and phosphorylated-AMPK (p-AMPK) levels, total CREB and phosphorylated-CREB (p-CREB) levels and PGC-1α performed on whole cell lysates from PS70 cells exposed to either vehicle, Insulin or Honokiol for 24 h. The graph displays the statistical analysis of the p-AMPK/AMPK and p-CREB/CREB ratio calculated by densitometric analysis of band intensity normalized to the corresponding β-Actin used as loading control; PGC-1α normalized to corresponding GAPDH level, used as loading control. Data, means ± SEM are expressed as percentage of vehicle-treated control; n = 3 under each condition. Significance was calculated with Student's t test, *p

    Journal: PLoS ONE

    Article Title: SIRT3 activator Honokiol attenuates β-Amyloid by modulating amyloidogenic pathway

    doi: 10.1371/journal.pone.0190350

    Figure Lengend Snippet: Effect of Honokiol on SIRT3 activation and AMPK-CREB-PGC1α pathway. Effect of increasing concentrations of insulin (0, 1, 5 and 10nM) on TsAβ (Total secreted Aβ) along with the densitometric analysis of band intensity normalized to β-Actin. (B) Representative Western blot of SIRT3 performed on whole cell lysates from PS70 cells exposed to either vehicle (DMSO) or 5 and 10μM Honokiol for 24 and 48 h. The graph displays the densitometric analysis of band intensity of the SIRT3 normalized to the corresponding GADPH level, used as loading control. (C) Effect of Insulin 10nM and Honokiol (5 and 10μM) on p-AMPK/AMPK ratio. Representative Western blot of total AMPK and phosphorylated-AMPK (p-AMPK) levels, total CREB and phosphorylated-CREB (p-CREB) levels and PGC-1α performed on whole cell lysates from PS70 cells exposed to either vehicle, Insulin or Honokiol for 24 h. The graph displays the statistical analysis of the p-AMPK/AMPK and p-CREB/CREB ratio calculated by densitometric analysis of band intensity normalized to the corresponding β-Actin used as loading control; PGC-1α normalized to corresponding GAPDH level, used as loading control. Data, means ± SEM are expressed as percentage of vehicle-treated control; n = 3 under each condition. Significance was calculated with Student's t test, *p

    Article Snippet: Primary antibodies used in this study included: AMPK (#2532), phospho-AMPK Thr172 (#2535), CREB (#4820) from CST; Anti-SIRT3 antibody (ab86671), Anti-PGC1α (ab54481), Anti-beta Amyloid 1–42 antibody (ab12267), Anti-beta Amyloid 1–40 antibody [BDI350] (ab20068), Anti-beta Actin antibody (ab8227), Anti-GAPDH (ab8245), Anti-ADAM10 antibody [EPR5622] (ab124695) from abcam; APP C-terminal antibody pAb751/770 (EMD Biosciences, La Jolla, CA, USA); Anti-BACE1 monoclonal antibody (MAB5308), anti-ADAM10 polyclonal antibody and Anti phospho-CREB (pAb06-519) from Merck Millipore; 6E10 (against sAPPα and β-CTF) and anti-sAPPβ antibodies from Covance.

    Techniques: Activation Assay, Western Blot, Pyrolysis Gas Chromatography

    Brain levels of PGC-1α protein ( a ) and mRNA ( b ). Representative western blot assays are included in lower part of a . Band of PGC-1α was located in second place at 100 kDa. Tubulin used as loading control. Expression levels of mRNA were normalized to PGK1 and B2M mRNA expression. Citrate synthase (CS) activity as a marker of mitochondrial content was determined in isolated brain mitochondria using a photometrical assay ( c ). Animals belonged to three different study groups (wild-type (control) , Thy-1 AβPP SL (control) , and treatment group Thy-1 AβPP SL (MH84) ). Data represent means ± SEM. N = 11 (six females, five males); one-way ANOVA with Tukey’s multiple comparison post test (* p

    Journal: Alzheimer's Research & Therapy

    Article Title: MH84 improves mitochondrial dysfunction in a mouse model of early Alzheimer’s disease

    doi: 10.1186/s13195-018-0342-6

    Figure Lengend Snippet: Brain levels of PGC-1α protein ( a ) and mRNA ( b ). Representative western blot assays are included in lower part of a . Band of PGC-1α was located in second place at 100 kDa. Tubulin used as loading control. Expression levels of mRNA were normalized to PGK1 and B2M mRNA expression. Citrate synthase (CS) activity as a marker of mitochondrial content was determined in isolated brain mitochondria using a photometrical assay ( c ). Animals belonged to three different study groups (wild-type (control) , Thy-1 AβPP SL (control) , and treatment group Thy-1 AβPP SL (MH84) ). Data represent means ± SEM. N = 11 (six females, five males); one-way ANOVA with Tukey’s multiple comparison post test (* p

    Article Snippet: To detect protein expression of PGC-1α in the brain, Anti-PGC-1α antibody (ab106814; abcam) was used.

    Techniques: Pyrolysis Gas Chromatography, Western Blot, Expressing, Activity Assay, Marker, Isolation

    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

    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

    Knockdown of PRL-3 decreases the expression of PGC-1α targeted SOD2 and UCP2. Notes: ( A ) Western blot (up) and RT-PCR (bottom) results showed that knockdown of RAP1 decreased the expression of PGC-1α. ( B ) Western blot (up) and RT-PCR (bottom) results showed that knockdown of PRL-3 decreased the expression of PGC-1α. ( C ) Knockdown of RAP1 in PRL-3 overexpressing cells suppressed the upregulation of PGC-1α. PRL-3 was overexpressed through transfection of pcDNA3-PRL-3-myc, and pcDNA3-vector-myc as control. Then RAP1 was knockdown through shRNA lentivirus infection. ( D ) Measurement of relative mtDNA by RT-PCR after knockdown of either PRL-3 or RAP1. ( E ) Knockdown of either RAP1 (up) or PRL-3 (bottom) decreased the mRNA level of SOD2 and UCP2. ( F ) Gene expression correlation analysis between PRL-3 and SOD2, RAP1 and SOD2 in TCGA data (PRL-3-SOD2: P =0.0012, R =0.49; RAP1-SOD2: P

    Journal: Cancer Management and Research

    Article Title: Knockdown of PRL-3 increases mitochondrial superoxide anion production through transcriptional regulation of RAP1

    doi: 10.2147/CMAR.S165344

    Figure Lengend Snippet: Knockdown of PRL-3 decreases the expression of PGC-1α targeted SOD2 and UCP2. Notes: ( A ) Western blot (up) and RT-PCR (bottom) results showed that knockdown of RAP1 decreased the expression of PGC-1α. ( B ) Western blot (up) and RT-PCR (bottom) results showed that knockdown of PRL-3 decreased the expression of PGC-1α. ( C ) Knockdown of RAP1 in PRL-3 overexpressing cells suppressed the upregulation of PGC-1α. PRL-3 was overexpressed through transfection of pcDNA3-PRL-3-myc, and pcDNA3-vector-myc as control. Then RAP1 was knockdown through shRNA lentivirus infection. ( D ) Measurement of relative mtDNA by RT-PCR after knockdown of either PRL-3 or RAP1. ( E ) Knockdown of either RAP1 (up) or PRL-3 (bottom) decreased the mRNA level of SOD2 and UCP2. ( F ) Gene expression correlation analysis between PRL-3 and SOD2, RAP1 and SOD2 in TCGA data (PRL-3-SOD2: P =0.0012, R =0.49; RAP1-SOD2: P

    Article Snippet: RAP1 (A300-306A-2, Bethyl Laboratories, Montgomery, TX, USA), GAPDH (ab8245, Abcam, Cambridge, UK), PGC-1α (#2178, Cell Signaling Technology, Danvers, MA, USA), and c-Myc (sc-40, Santa Cruz Biotechnology, Dallas, TX, USA).

    Techniques: Expressing, Pyrolysis Gas Chromatography, Western Blot, Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation, shRNA, Infection

    A Schematic depiction of the mechanism of CD105/BMP signaling in conferring radiation resistance. Radiation of prostate cancer results in upregulation of cell surface CD105 expression. The ensuing CD105/BMP signaling is essential and necessary for SIRT1 expression and downstream activity. SIRT1 facilitates DNA damage repair, destabilizes p53, and activates of PGC-1α in response to radiation. Consequently, CD105 can regulate glycolysis and mitochondrial biogenesis to meet the higher energy demands necessary for survival following irradiation. The loss of functional p53 enables glycolysis as a source of ATP generation and survival

    Journal: Oncogene

    Article Title: Antagonizing CD105 enhances radiation sensitivity in prostate cancer

    doi: 10.1038/s41388-018-0278-0

    Figure Lengend Snippet: A Schematic depiction of the mechanism of CD105/BMP signaling in conferring radiation resistance. Radiation of prostate cancer results in upregulation of cell surface CD105 expression. The ensuing CD105/BMP signaling is essential and necessary for SIRT1 expression and downstream activity. SIRT1 facilitates DNA damage repair, destabilizes p53, and activates of PGC-1α in response to radiation. Consequently, CD105 can regulate glycolysis and mitochondrial biogenesis to meet the higher energy demands necessary for survival following irradiation. The loss of functional p53 enables glycolysis as a source of ATP generation and survival

    Article Snippet: Whole-lysate western blots were probed for the following antibodies phos-SMAD1/5 (9516, Cell Signaling Technologies), SIRT1 (9475, Cell Signaling Technologies), p21 (4060, Cell Signaling Technologies), PGC-1α (ST1202, EMD Millipore), Total OXPHOS Rodent (ab110413, Abcam), PGC-1α (ST1202, EMD Millipore), lamin B (sc-6217, Santa Cruz Biotechnology), Rho A (sc-418, Santa Cruz Biotechnology), p53 (sc-126, Santa Cruz Biotechnology), K382 acetyl-p53 (2525, Cell Signaling Technologies), and β-actin (sc-47778, Santa Cruz Biotechnology).

    Techniques: Expressing, Activity Assay, Pyrolysis Gas Chromatography, Irradiation, Functional Assay

    PGC-1α and mitochondrial biogenesis are regulated by CD105/BMP. 22Rv1 cells were incubated with IgG or TRC105 with or without 4 Gy irradiation. All measurements were made 72 h post irradiation. a Western blot for whole-cell lysate, nuclear, and cytoplasmic fractions were independently analyzed for PGC-1α expression. Loading controls included β-actin (whole cell), lamin B (nuclear marker), and Rho A (cytoplasm marker). Molecular weights (kDa) of the ladder are indicated. b Immunofluorescent localization of PGC-1α (green) was visualized with DAPI (blue) nuclear counterstain. c The mRNA expression of PGC-1α target genes, NRF1 , MTFA , and CPT1C were measured. MRNA expression was normalized to GAPDH and untreated. d Mitochondrial DNA (mtDNA) was measured from total DNA extracts and normalized to nuclear DNA and to untreated. Data are reported as means ± S.D. of three independent experiments (** p

    Journal: Oncogene

    Article Title: Antagonizing CD105 enhances radiation sensitivity in prostate cancer

    doi: 10.1038/s41388-018-0278-0

    Figure Lengend Snippet: PGC-1α and mitochondrial biogenesis are regulated by CD105/BMP. 22Rv1 cells were incubated with IgG or TRC105 with or without 4 Gy irradiation. All measurements were made 72 h post irradiation. a Western blot for whole-cell lysate, nuclear, and cytoplasmic fractions were independently analyzed for PGC-1α expression. Loading controls included β-actin (whole cell), lamin B (nuclear marker), and Rho A (cytoplasm marker). Molecular weights (kDa) of the ladder are indicated. b Immunofluorescent localization of PGC-1α (green) was visualized with DAPI (blue) nuclear counterstain. c The mRNA expression of PGC-1α target genes, NRF1 , MTFA , and CPT1C were measured. MRNA expression was normalized to GAPDH and untreated. d Mitochondrial DNA (mtDNA) was measured from total DNA extracts and normalized to nuclear DNA and to untreated. Data are reported as means ± S.D. of three independent experiments (** p

    Article Snippet: Whole-lysate western blots were probed for the following antibodies phos-SMAD1/5 (9516, Cell Signaling Technologies), SIRT1 (9475, Cell Signaling Technologies), p21 (4060, Cell Signaling Technologies), PGC-1α (ST1202, EMD Millipore), Total OXPHOS Rodent (ab110413, Abcam), PGC-1α (ST1202, EMD Millipore), lamin B (sc-6217, Santa Cruz Biotechnology), Rho A (sc-418, Santa Cruz Biotechnology), p53 (sc-126, Santa Cruz Biotechnology), K382 acetyl-p53 (2525, Cell Signaling Technologies), and β-actin (sc-47778, Santa Cruz Biotechnology).

    Techniques: Pyrolysis Gas Chromatography, Incubation, Irradiation, Western Blot, Expressing, Marker

    The effect of BBR addition in the preservation solution before ischemia on the gene expression of Sirt 1 and 3, PGC-1α, MnSOD2, COX IV and NDUFS8 as evaluated by qPCR. Tissue samples were obtained from animals sacrificed after 12 h of hepatic exposure to preservation solution and 1 h of reperfusion, as described in the Materials and Methods Section. Control animals were sham operated. I/R livers (Group B) had a reduced gene expression of SirT1, the nuclearly-encoded mitochondrial respiratory complex IV subunit IV (COX IV) and PGC-1α, which were prevented by BBR pre-treatment (Group A). No differences were found in the gene expression of SirT3, the mitochondrial antioxidant protein MnSOD2 and the mitochondrially-encoded mitochondrial respiratory complex I iron-sulfur protein 8 (NDUFS8). Boxes represent the means ± 5–95 percentile of an n = 6 different experiments. * indicates a statistically-significant difference ( p

    Journal: International Journal of Molecular Sciences

    Article Title: Addition of Berberine to Preservation Solution in an Animal Model of Ex Vivo Liver Transplant Preserves Mitochondrial Function and Bioenergetics from the Damage Induced by Ischemia/Reperfusion

    doi: 10.3390/ijms19010284

    Figure Lengend Snippet: The effect of BBR addition in the preservation solution before ischemia on the gene expression of Sirt 1 and 3, PGC-1α, MnSOD2, COX IV and NDUFS8 as evaluated by qPCR. Tissue samples were obtained from animals sacrificed after 12 h of hepatic exposure to preservation solution and 1 h of reperfusion, as described in the Materials and Methods Section. Control animals were sham operated. I/R livers (Group B) had a reduced gene expression of SirT1, the nuclearly-encoded mitochondrial respiratory complex IV subunit IV (COX IV) and PGC-1α, which were prevented by BBR pre-treatment (Group A). No differences were found in the gene expression of SirT3, the mitochondrial antioxidant protein MnSOD2 and the mitochondrially-encoded mitochondrial respiratory complex I iron-sulfur protein 8 (NDUFS8). Boxes represent the means ± 5–95 percentile of an n = 6 different experiments. * indicates a statistically-significant difference ( p

    Article Snippet: Membranes were blocked with 5% blocking solution (Bio-Rad) for 2 h and incubated in Tris-buffered saline (TBS) supplemented with 1% Tween-20 (TBS-T) and blocking solution 0.5%, overnight at 4 °C, with anti-PGC-1α (Cell Signaling Technology, Beverly, MA, USA, 1:100), anti-SirT1 (Cell Signaling Technology, 1:1000), anti-SirT3 (Cell Signaling Technology, 1:500), anti-LC3 (Sigma-Aldrich, 1:1000) or anti-β-actin (Sigma-Aldrich, 1:5000) antibodies.

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

    The effect of BBR addition in the preservation solution before ischemia on the content of LC3, Sirt 1 and 3, PGC-1α, as evaluated by Western blot. Tissue samples were obtained from animals sacrificed after 12 h of hepatic exposure to preservation solution and 1 h of reperfusion, as described in the Materials and Methods Section. Control animals were sham operated. I/R livers (Group B) had a reduced content in SirT1, SirT3 and PGC-1α (all these effects were prevented by BBR addition; Group A). The active form of LC3 (LC3 II) was elevated in Group A (BBR addition), when compared to Group B (I/R). The image shows a representative Western blot of these experiments for the respective proteins, from six independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Addition of Berberine to Preservation Solution in an Animal Model of Ex Vivo Liver Transplant Preserves Mitochondrial Function and Bioenergetics from the Damage Induced by Ischemia/Reperfusion

    doi: 10.3390/ijms19010284

    Figure Lengend Snippet: The effect of BBR addition in the preservation solution before ischemia on the content of LC3, Sirt 1 and 3, PGC-1α, as evaluated by Western blot. Tissue samples were obtained from animals sacrificed after 12 h of hepatic exposure to preservation solution and 1 h of reperfusion, as described in the Materials and Methods Section. Control animals were sham operated. I/R livers (Group B) had a reduced content in SirT1, SirT3 and PGC-1α (all these effects were prevented by BBR addition; Group A). The active form of LC3 (LC3 II) was elevated in Group A (BBR addition), when compared to Group B (I/R). The image shows a representative Western blot of these experiments for the respective proteins, from six independent experiments.

    Article Snippet: Membranes were blocked with 5% blocking solution (Bio-Rad) for 2 h and incubated in Tris-buffered saline (TBS) supplemented with 1% Tween-20 (TBS-T) and blocking solution 0.5%, overnight at 4 °C, with anti-PGC-1α (Cell Signaling Technology, Beverly, MA, USA, 1:100), anti-SirT1 (Cell Signaling Technology, 1:1000), anti-SirT3 (Cell Signaling Technology, 1:500), anti-LC3 (Sigma-Aldrich, 1:1000) or anti-β-actin (Sigma-Aldrich, 1:5000) antibodies.

    Techniques: Preserving, Pyrolysis Gas Chromatography, Western Blot

    METRNL alleviates inflammation and insulin resistance through a PPARδ-mediated pathway. a Western blot analysis of PPARδ expression in differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. b Confirmation of PPARδ siRNA efficiency in differentiated C2C12 cells. Western blot analysis of palmitate (200 μM)-induced inflammation markers in PPARα siRNA (20 nM)-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. c Western blot analysis of the palmitate-induced impairment of IRS-1 and Akt phosphorylation in PPARδ siRNA-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. Human insulin (10 nM) was used to stimulate insulin signaling for 3 min. d Western blot analysis of PPARδ expression in soleus muscle of mice treated with HFD and METRNL (5 animals/treatment group). e Western blot analysis of METRNL (200 ng/mL)-induced PGC1α expression in PPARδ siRNA (20 nM)-transfected differentiated C2C12 cells for 24 h. Western blot analysis of f AMPK phosphorylation in PPARδ siRNA or g PPARδ expression in AMPK siRNA-transfected differentiated C2C12 cells treated with METRNL (200 ng/mL) for 24 h. Confirmation of PPARδ siRNA efficiency in skeletal muscle of mice. Western blot analysis of IRS-1 and Akt phosphorylation ( h ), and inflammatory markers ( i ) in PPARδ siRNA-transfected skeletal muscle of experimental mice. j Western blot analysis of PPARδ expression in AMPK siRNA-transfected skeletal muscle of experimental mice. k Western blot analysis of AMPK phosphorylation in PPARδ siRNA-transfected skeletal muscle of experimental mice. Means ± SEM were obtained from three separate experiments or five animals. *** P

    Journal: Experimental & Molecular Medicine

    Article Title: METRNL attenuates lipid-induced inflammation and insulin resistance via AMPK or PPARδ-dependent pathways in skeletal muscle of mice

    doi: 10.1038/s12276-018-0147-5

    Figure Lengend Snippet: METRNL alleviates inflammation and insulin resistance through a PPARδ-mediated pathway. a Western blot analysis of PPARδ expression in differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. b Confirmation of PPARδ siRNA efficiency in differentiated C2C12 cells. Western blot analysis of palmitate (200 μM)-induced inflammation markers in PPARα siRNA (20 nM)-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. c Western blot analysis of the palmitate-induced impairment of IRS-1 and Akt phosphorylation in PPARδ siRNA-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. Human insulin (10 nM) was used to stimulate insulin signaling for 3 min. d Western blot analysis of PPARδ expression in soleus muscle of mice treated with HFD and METRNL (5 animals/treatment group). e Western blot analysis of METRNL (200 ng/mL)-induced PGC1α expression in PPARδ siRNA (20 nM)-transfected differentiated C2C12 cells for 24 h. Western blot analysis of f AMPK phosphorylation in PPARδ siRNA or g PPARδ expression in AMPK siRNA-transfected differentiated C2C12 cells treated with METRNL (200 ng/mL) for 24 h. Confirmation of PPARδ siRNA efficiency in skeletal muscle of mice. Western blot analysis of IRS-1 and Akt phosphorylation ( h ), and inflammatory markers ( i ) in PPARδ siRNA-transfected skeletal muscle of experimental mice. j Western blot analysis of PPARδ expression in AMPK siRNA-transfected skeletal muscle of experimental mice. k Western blot analysis of AMPK phosphorylation in PPARδ siRNA-transfected skeletal muscle of experimental mice. Means ± SEM were obtained from three separate experiments or five animals. *** P

    Article Snippet: Anti-insulin receptor substrate (IRS)-1 (1:2500), anti-phospho-Akt (Ser473; 1:1000), anti-Akt (1:1000), anti-phospho-AMPK (Thr172; 1:1000), anti-AMPK (1:2500), anti-NFκBp65 (1:2500), anti-phospho-IκBα (Ser32; 1:1000), anti-phospho-eIF2α (Ser51; 1:1000), anti-eIF2α (1:1000), anti-PPARδ (1:2500), anti-PPARα (1:1000), anti-PPARγ (1:1000), anti-phospho-LKB1 (Ser428; 1:1000), anti-LKB1 (1:2500), anti-phospho-acetyl-CoA carboxylase (ACC) (Ser79; 1:1000), anti-ACC (1:2500), and anti-PGC1α (1:1000) antibodies were procured from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Western Blot, Expressing, Transfection, Mouse Assay

    METRNL ameliorates inflammation and insulin resistance via an AMPK-mediated pathway. a Western blot analysis of LKB1, AMPK, and ACC phosphorylation in differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. b Confirmation of AMPK siRNA efficiency in differentiated C2C12 cells. Western blot analysis of palmitate (200 μM)-induced inflammation markers in AMPK siRNA (20 nM)-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. c Western blot analysis of the palmitate-induced impairment of IRS-1 and Akt phosphorylation in AMPK siRNA-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. Human insulin (10 nM) was used to stimulate insulin signaling for 3 min. d Western blot analysis of AMPK phosphorylation in soleus muscle of mice treated with HFD and METRNL (5 animals/treatment group). e Western blot analysis of PGC1α expression in differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. f Western blot analysis of METRNL (200 ng/mL)-induced PGC1α expression in AMPK siRNA (20 nM)-transfected differentiated C2C12 cells for 24 h. Confirmation of AMPK siRNA efficiency in skeletal muscle of mice. Western blot analysis of IRS-1 and Akt phosphorylation ( g ), and inflammatory markers ( h ) in AMPK siRNA-transfected skeletal muscle of experimental mice. Means ± SEM were obtained from three separate experiments or five animals. *** P

    Journal: Experimental & Molecular Medicine

    Article Title: METRNL attenuates lipid-induced inflammation and insulin resistance via AMPK or PPARδ-dependent pathways in skeletal muscle of mice

    doi: 10.1038/s12276-018-0147-5

    Figure Lengend Snippet: METRNL ameliorates inflammation and insulin resistance via an AMPK-mediated pathway. a Western blot analysis of LKB1, AMPK, and ACC phosphorylation in differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. b Confirmation of AMPK siRNA efficiency in differentiated C2C12 cells. Western blot analysis of palmitate (200 μM)-induced inflammation markers in AMPK siRNA (20 nM)-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. c Western blot analysis of the palmitate-induced impairment of IRS-1 and Akt phosphorylation in AMPK siRNA-transfected differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. Human insulin (10 nM) was used to stimulate insulin signaling for 3 min. d Western blot analysis of AMPK phosphorylation in soleus muscle of mice treated with HFD and METRNL (5 animals/treatment group). e Western blot analysis of PGC1α expression in differentiated C2C12 cells treated with METRNL (0–200 ng/mL) for 24 h. f Western blot analysis of METRNL (200 ng/mL)-induced PGC1α expression in AMPK siRNA (20 nM)-transfected differentiated C2C12 cells for 24 h. Confirmation of AMPK siRNA efficiency in skeletal muscle of mice. Western blot analysis of IRS-1 and Akt phosphorylation ( g ), and inflammatory markers ( h ) in AMPK siRNA-transfected skeletal muscle of experimental mice. Means ± SEM were obtained from three separate experiments or five animals. *** P

    Article Snippet: Anti-insulin receptor substrate (IRS)-1 (1:2500), anti-phospho-Akt (Ser473; 1:1000), anti-Akt (1:1000), anti-phospho-AMPK (Thr172; 1:1000), anti-AMPK (1:2500), anti-NFκBp65 (1:2500), anti-phospho-IκBα (Ser32; 1:1000), anti-phospho-eIF2α (Ser51; 1:1000), anti-eIF2α (1:1000), anti-PPARδ (1:2500), anti-PPARα (1:1000), anti-PPARγ (1:1000), anti-phospho-LKB1 (Ser428; 1:1000), anti-LKB1 (1:2500), anti-phospho-acetyl-CoA carboxylase (ACC) (Ser79; 1:1000), anti-ACC (1:2500), and anti-PGC1α (1:1000) antibodies were procured from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Western Blot, Transfection, Mouse Assay, Expressing

    PGC-1α knockdown inhibits T3-induced dendritic growth in Purkinje cells. (A) HEK293T cells expressing mouse PGC-1α were transfected with PGC-1α-scramble (scr) or short hairpin RNA (shRNA) construct and analyzed by western blotting with anti-PGC-1α and anti-β-actin antibodies. (B,C) Cultured Purkinje cells were transfected with PGC-1α-shRNA or scr-shRNA (control) constructs and stained for Calbindin and PGC-1α (B) or Calbindin and cytochrome C oxidase IV (COX-IV) (C) at 10 DIV. Scale bars, 10 μm. (D) Representative images of Purkinje cells transfected with scr shRNA (control), PGC-1α shRNA (PGC-1α shRNA), or PGC-1α shRNA plus an shRNA-resistant mutant of PGC-1α (shRNA + rescue). Cells were cultured in the presence of 10 nM T3 until 10 DIV and immunostained with anti-Calbindin and anti-PDH antibodies. Boxed regions in the upper panels are enlarged in lower panels. Scale bars, 20 μm. (E–G) Quantitative analyses of the total dendritic length (E) , number of dendritic branches (F) and PDH signal (G) . Data represent mean ± SEM, N = 30 for each point, ** p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Thyroid Hormone Induces PGC-1α during Dendritic Outgrowth in Mouse Cerebellar Purkinje Cells

    doi: 10.3389/fncel.2017.00133

    Figure Lengend Snippet: PGC-1α knockdown inhibits T3-induced dendritic growth in Purkinje cells. (A) HEK293T cells expressing mouse PGC-1α were transfected with PGC-1α-scramble (scr) or short hairpin RNA (shRNA) construct and analyzed by western blotting with anti-PGC-1α and anti-β-actin antibodies. (B,C) Cultured Purkinje cells were transfected with PGC-1α-shRNA or scr-shRNA (control) constructs and stained for Calbindin and PGC-1α (B) or Calbindin and cytochrome C oxidase IV (COX-IV) (C) at 10 DIV. Scale bars, 10 μm. (D) Representative images of Purkinje cells transfected with scr shRNA (control), PGC-1α shRNA (PGC-1α shRNA), or PGC-1α shRNA plus an shRNA-resistant mutant of PGC-1α (shRNA + rescue). Cells were cultured in the presence of 10 nM T3 until 10 DIV and immunostained with anti-Calbindin and anti-PDH antibodies. Boxed regions in the upper panels are enlarged in lower panels. Scale bars, 20 μm. (E–G) Quantitative analyses of the total dendritic length (E) , number of dendritic branches (F) and PDH signal (G) . Data represent mean ± SEM, N = 30 for each point, ** p

    Article Snippet: Antibodies used for immunofluorescence were as follows: mouse anti-Calbindin D28K (Swant): rabbit anti-Calbindin D28K (Millipore): goat anti-Calbindin (Santa Cruz): mouse anti-Pyruvate dehydrogenase (PDH; Abcam): rabbit anti- PGC-1α (Abcam): rabbit anti-DsRed (Clontech): mouse anti-Pax-6 (R & D): rabbit anti-cytochrome C oxidase IV (COX-IV; Abcam); and Alexa 488-, Alexa 568-, or Alexa 647-conjugated anti-mouse, anti-rabbit, and anti-goat IgG (Invitrogen).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Transfection, shRNA, Construct, Western Blot, Cell Culture, Staining, Mutagenesis

    PGC-1α overexpression enhances dendritic outgrowth of Purkinje cells in the absence of T3. (A,B) The morphology of Purkinje cells transfected with tdTomato (control) or PGC-1α-mCherry (+PGC-1α). Cells were cultured with (B) or without (A) T3 and stained for Calbindin at 10 DIV. Scale bars, 20 μm. PGC-1α overexpression induced dendritic outgrowth in the absence of T3 (A) , but not in the presence of T3 (B) . (C–E) Quantitative analyses of total dendritic length (C) branch numbers (D) and PDH signal (E) . Purkinje cells expressing tdTomato (control) or PGC-1α were cultured with or without T3 treatment. N = 30 for all data points. Data represent mean ± SEM, *** p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Thyroid Hormone Induces PGC-1α during Dendritic Outgrowth in Mouse Cerebellar Purkinje Cells

    doi: 10.3389/fncel.2017.00133

    Figure Lengend Snippet: PGC-1α overexpression enhances dendritic outgrowth of Purkinje cells in the absence of T3. (A,B) The morphology of Purkinje cells transfected with tdTomato (control) or PGC-1α-mCherry (+PGC-1α). Cells were cultured with (B) or without (A) T3 and stained for Calbindin at 10 DIV. Scale bars, 20 μm. PGC-1α overexpression induced dendritic outgrowth in the absence of T3 (A) , but not in the presence of T3 (B) . (C–E) Quantitative analyses of total dendritic length (C) branch numbers (D) and PDH signal (E) . Purkinje cells expressing tdTomato (control) or PGC-1α were cultured with or without T3 treatment. N = 30 for all data points. Data represent mean ± SEM, *** p

    Article Snippet: Antibodies used for immunofluorescence were as follows: mouse anti-Calbindin D28K (Swant): rabbit anti-Calbindin D28K (Millipore): goat anti-Calbindin (Santa Cruz): mouse anti-Pyruvate dehydrogenase (PDH; Abcam): rabbit anti- PGC-1α (Abcam): rabbit anti-DsRed (Clontech): mouse anti-Pax-6 (R & D): rabbit anti-cytochrome C oxidase IV (COX-IV; Abcam); and Alexa 488-, Alexa 568-, or Alexa 647-conjugated anti-mouse, anti-rabbit, and anti-goat IgG (Invitrogen).

    Techniques: Pyrolysis Gas Chromatography, Over Expression, Transfection, Cell Culture, Staining, Expressing

    Knockdown of PGC-1α inhibits dendritic outgrowth in vivo Purkinje cells. (A) Representative images of Purkinje cells transfected with scr shRNA (control) or PGC-1α shRNA construct. Scale bar, 20 μm. (B) Dual color images of GFP derived from shRNA constructs (green) and immunostaining with anti-PGC-1α (magenta). Scale bar, 20 μm. (C,D) Quantitative analyses of the total dendritic length (C) and number of dendritic branches (D) in Purkinje cells expressing scr shRNA (control) or PGC-1α shRNA constructs. Data represent mean ± SEM, N = 12 cells from three mice, *** p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Thyroid Hormone Induces PGC-1α during Dendritic Outgrowth in Mouse Cerebellar Purkinje Cells

    doi: 10.3389/fncel.2017.00133

    Figure Lengend Snippet: Knockdown of PGC-1α inhibits dendritic outgrowth in vivo Purkinje cells. (A) Representative images of Purkinje cells transfected with scr shRNA (control) or PGC-1α shRNA construct. Scale bar, 20 μm. (B) Dual color images of GFP derived from shRNA constructs (green) and immunostaining with anti-PGC-1α (magenta). Scale bar, 20 μm. (C,D) Quantitative analyses of the total dendritic length (C) and number of dendritic branches (D) in Purkinje cells expressing scr shRNA (control) or PGC-1α shRNA constructs. Data represent mean ± SEM, N = 12 cells from three mice, *** p

    Article Snippet: Antibodies used for immunofluorescence were as follows: mouse anti-Calbindin D28K (Swant): rabbit anti-Calbindin D28K (Millipore): goat anti-Calbindin (Santa Cruz): mouse anti-Pyruvate dehydrogenase (PDH; Abcam): rabbit anti- PGC-1α (Abcam): rabbit anti-DsRed (Clontech): mouse anti-Pax-6 (R & D): rabbit anti-cytochrome C oxidase IV (COX-IV; Abcam); and Alexa 488-, Alexa 568-, or Alexa 647-conjugated anti-mouse, anti-rabbit, and anti-goat IgG (Invitrogen).

    Techniques: Pyrolysis Gas Chromatography, In Vivo, Transfection, shRNA, Construct, Derivative Assay, Immunostaining, Expressing, Mouse Assay

    PGC-1α expression in the developing cerebellar cortex. (A) Shape changes of Purkinje cell dendrites during postnatal development. (B,C) Sagittal cerebellar sections were immunostained for PGC-1α and Calbindin at different ages of development and were observed at low (B) and high (C) magnification. Scale bars, 40 μm (B) and 20 μm (C) . PGC-1α was predominantly detected in the Purkinje cells from P7. EGL, external granule layer; ML, molecular layer; PCL, Purkinje cell layer; IGL, internal granule layer; WM, white matter.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Thyroid Hormone Induces PGC-1α during Dendritic Outgrowth in Mouse Cerebellar Purkinje Cells

    doi: 10.3389/fncel.2017.00133

    Figure Lengend Snippet: PGC-1α expression in the developing cerebellar cortex. (A) Shape changes of Purkinje cell dendrites during postnatal development. (B,C) Sagittal cerebellar sections were immunostained for PGC-1α and Calbindin at different ages of development and were observed at low (B) and high (C) magnification. Scale bars, 40 μm (B) and 20 μm (C) . PGC-1α was predominantly detected in the Purkinje cells from P7. EGL, external granule layer; ML, molecular layer; PCL, Purkinje cell layer; IGL, internal granule layer; WM, white matter.

    Article Snippet: Antibodies used for immunofluorescence were as follows: mouse anti-Calbindin D28K (Swant): rabbit anti-Calbindin D28K (Millipore): goat anti-Calbindin (Santa Cruz): mouse anti-Pyruvate dehydrogenase (PDH; Abcam): rabbit anti- PGC-1α (Abcam): rabbit anti-DsRed (Clontech): mouse anti-Pax-6 (R & D): rabbit anti-cytochrome C oxidase IV (COX-IV; Abcam); and Alexa 488-, Alexa 568-, or Alexa 647-conjugated anti-mouse, anti-rabbit, and anti-goat IgG (Invitrogen).

    Techniques: Pyrolysis Gas Chromatography, Expressing

    PGC-1α expression in Purkinje cells is downregulated in hypothyroid mice. (A–C) PGC-1α expression was compared at P7 (A) , P9 (B) and P14 (C) . Representative images from four mice in each condition are shown. Dendritic growth of Purkinje cells is retarded in the hypothyroid condition. At P14 (C) , the EGL is thicker in hypothyroid animals compared to control animals. Each section was immunostained with anti-Calbindin and anti-PGC-1α. DNA was counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bars, 40 μm. (D) Quantitative comparison of PGC-1α expression in Purkinje cells in control (euthyroid) and hypothyroid animals. Data represent mean ± SEM, N = 15 cells from four mice from two independent experiments for each points, * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Thyroid Hormone Induces PGC-1α during Dendritic Outgrowth in Mouse Cerebellar Purkinje Cells

    doi: 10.3389/fncel.2017.00133

    Figure Lengend Snippet: PGC-1α expression in Purkinje cells is downregulated in hypothyroid mice. (A–C) PGC-1α expression was compared at P7 (A) , P9 (B) and P14 (C) . Representative images from four mice in each condition are shown. Dendritic growth of Purkinje cells is retarded in the hypothyroid condition. At P14 (C) , the EGL is thicker in hypothyroid animals compared to control animals. Each section was immunostained with anti-Calbindin and anti-PGC-1α. DNA was counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bars, 40 μm. (D) Quantitative comparison of PGC-1α expression in Purkinje cells in control (euthyroid) and hypothyroid animals. Data represent mean ± SEM, N = 15 cells from four mice from two independent experiments for each points, * p

    Article Snippet: Antibodies used for immunofluorescence were as follows: mouse anti-Calbindin D28K (Swant): rabbit anti-Calbindin D28K (Millipore): goat anti-Calbindin (Santa Cruz): mouse anti-Pyruvate dehydrogenase (PDH; Abcam): rabbit anti- PGC-1α (Abcam): rabbit anti-DsRed (Clontech): mouse anti-Pax-6 (R & D): rabbit anti-cytochrome C oxidase IV (COX-IV; Abcam); and Alexa 488-, Alexa 568-, or Alexa 647-conjugated anti-mouse, anti-rabbit, and anti-goat IgG (Invitrogen).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Mouse Assay

    PGC-1α expression is induced in cultured Purkinje cells by T3 treatment. (A) Cerebellar cells were cultured in the presence of 10 nM T3 and then immunostained with anti-Calbindin and PGC-1α at the indicated day in culture. Boxed regions in the upper panels are enlarged in lower panels. PGC-1α expression is gradually increased in Purkinje cells (arrows) from 6 DIV. Scale bars, 20 μm. (B) Quantitative comparison of PGC-1α expression in Purkinje cells cultured with (black dots and line) or without (gray dots and line) T3. Data represent mean ± SEM, N = 15 cells for each points, *** p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Thyroid Hormone Induces PGC-1α during Dendritic Outgrowth in Mouse Cerebellar Purkinje Cells

    doi: 10.3389/fncel.2017.00133

    Figure Lengend Snippet: PGC-1α expression is induced in cultured Purkinje cells by T3 treatment. (A) Cerebellar cells were cultured in the presence of 10 nM T3 and then immunostained with anti-Calbindin and PGC-1α at the indicated day in culture. Boxed regions in the upper panels are enlarged in lower panels. PGC-1α expression is gradually increased in Purkinje cells (arrows) from 6 DIV. Scale bars, 20 μm. (B) Quantitative comparison of PGC-1α expression in Purkinje cells cultured with (black dots and line) or without (gray dots and line) T3. Data represent mean ± SEM, N = 15 cells for each points, *** p

    Article Snippet: Antibodies used for immunofluorescence were as follows: mouse anti-Calbindin D28K (Swant): rabbit anti-Calbindin D28K (Millipore): goat anti-Calbindin (Santa Cruz): mouse anti-Pyruvate dehydrogenase (PDH; Abcam): rabbit anti- PGC-1α (Abcam): rabbit anti-DsRed (Clontech): mouse anti-Pax-6 (R & D): rabbit anti-cytochrome C oxidase IV (COX-IV; Abcam); and Alexa 488-, Alexa 568-, or Alexa 647-conjugated anti-mouse, anti-rabbit, and anti-goat IgG (Invitrogen).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Cell Culture

    Molecular perturbation of PGC-1α inhibits dendritic outgrowth and mitochondrial activity in Purkinje cells. (A) Representative images of Purkinje cells overexpressing EGFP (control), EGFP-NRF1DN or FLAG-RIP140. Cells were stained for Calbindin at 10 DIV. Scale bar, 20 μm. (B–D) Quantitative analyses of the total dendritic length (B) , number of dendritic branches (C) and PDH signal (D) . N = 40 cells for control, 30 cells for NRF1DN and 30 cells for RIP140. Data represent mean ± SEM, ** p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Thyroid Hormone Induces PGC-1α during Dendritic Outgrowth in Mouse Cerebellar Purkinje Cells

    doi: 10.3389/fncel.2017.00133

    Figure Lengend Snippet: Molecular perturbation of PGC-1α inhibits dendritic outgrowth and mitochondrial activity in Purkinje cells. (A) Representative images of Purkinje cells overexpressing EGFP (control), EGFP-NRF1DN or FLAG-RIP140. Cells were stained for Calbindin at 10 DIV. Scale bar, 20 μm. (B–D) Quantitative analyses of the total dendritic length (B) , number of dendritic branches (C) and PDH signal (D) . N = 40 cells for control, 30 cells for NRF1DN and 30 cells for RIP140. Data represent mean ± SEM, ** p

    Article Snippet: Antibodies used for immunofluorescence were as follows: mouse anti-Calbindin D28K (Swant): rabbit anti-Calbindin D28K (Millipore): goat anti-Calbindin (Santa Cruz): mouse anti-Pyruvate dehydrogenase (PDH; Abcam): rabbit anti- PGC-1α (Abcam): rabbit anti-DsRed (Clontech): mouse anti-Pax-6 (R & D): rabbit anti-cytochrome C oxidase IV (COX-IV; Abcam); and Alexa 488-, Alexa 568-, or Alexa 647-conjugated anti-mouse, anti-rabbit, and anti-goat IgG (Invitrogen).

    Techniques: Pyrolysis Gas Chromatography, Activity Assay, Staining

    Alpha-synuclein binds to PGC1alpha promoter sequences in vitro and in vivo and in PD brain tissues

    Journal: Free radical biology & medicine

    Article Title: Selective binding of nuclear alpha-synuclein to the PGC1alpha promoter under conditions of oxidative stress may contribute to losses in mitochondrial function: implications for Parkinson's disease

    doi: 10.1016/j.freeradbiomed.2012.05.024

    Figure Lengend Snippet: Alpha-synuclein binds to PGC1alpha promoter sequences in vitro and in vivo and in PD brain tissues

    Article Snippet: Membranes were blocked with 5% powdered milk solution in 0.3% Triton/phosphate-buffered saline solution before incubation with either 1:500 alpha-synuclein (BD Transduction, San Jose CA), 1:250 PGC1alpha (Abcam, Cambridge, UK), or 1:1000 histone H3 or acetylated H3 (Abcam, Cambridge, UK) antibodies.

    Techniques: In Vitro, In Vivo

    Alpha synuclein overexpression results in reduced PGC1alpha promoter activity, reduced expression of PGC1alpha, and downstream transcriptional effects

    Journal: Free radical biology & medicine

    Article Title: Selective binding of nuclear alpha-synuclein to the PGC1alpha promoter under conditions of oxidative stress may contribute to losses in mitochondrial function: implications for Parkinson's disease

    doi: 10.1016/j.freeradbiomed.2012.05.024

    Figure Lengend Snippet: Alpha synuclein overexpression results in reduced PGC1alpha promoter activity, reduced expression of PGC1alpha, and downstream transcriptional effects

    Article Snippet: Membranes were blocked with 5% powdered milk solution in 0.3% Triton/phosphate-buffered saline solution before incubation with either 1:500 alpha-synuclein (BD Transduction, San Jose CA), 1:250 PGC1alpha (Abcam, Cambridge, UK), or 1:1000 histone H3 or acetylated H3 (Abcam, Cambridge, UK) antibodies.

    Techniques: Over Expression, Activity Assay, Expressing

    ( A ) Representative Western blot of p-p65 (Ser536), p65, p-STAT3 (Tyr705) and STAT3 in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). GAPDH was used as the loading control. ( B ) Representative Western blot of p65 and PGC-1α in the PGC-1α immunoprecipitate of the livers of the sham PGC-1α +/+ (WT) mice and at 1 h after cerulein-induced AP mice. ( C ) Representative Western blot of p-STAT3 (Tyr705) and p65 in the p65 immunoprecipitate of the livers of the PGC-1α +/+ (WT) mice and PGC-1α -/- (KO) mice with pancreatitis (Cerulein). IgG was used as the loading control. There were six mice per group.

    Journal: Antioxidants

    Article Title: Impairment of PGC-1 Alpha Up-Regulation Enhances Nitrosative Stress in the Liver during Acute Pancreatitis in Obese Mice

    doi: 10.3390/antiox9090887

    Figure Lengend Snippet: ( A ) Representative Western blot of p-p65 (Ser536), p65, p-STAT3 (Tyr705) and STAT3 in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). GAPDH was used as the loading control. ( B ) Representative Western blot of p65 and PGC-1α in the PGC-1α immunoprecipitate of the livers of the sham PGC-1α +/+ (WT) mice and at 1 h after cerulein-induced AP mice. ( C ) Representative Western blot of p-STAT3 (Tyr705) and p65 in the p65 immunoprecipitate of the livers of the PGC-1α +/+ (WT) mice and PGC-1α -/- (KO) mice with pancreatitis (Cerulein). IgG was used as the loading control. There were six mice per group.

    Article Snippet: The employed antibodies were: anti-β-tubulin (1:1000, ab6046 from Abcam, Cambridge, UK); anti-PGC-1α (1:500, sc-518025 from Santa Cruz Biotechnology, Dallas, TX, USA); anti-p65 (1:1000, #8242 from Cell Signaling Technology, Danvers, MA, USA); anti-phospho-p65 (Ser 536) (1:100, #3033 from Cell Signaling); anti-Nitro-tyrosine (1:1000, #9691 from Cell Signaling); anti-STAT3 (1:1000, #9132 from Cell Signaling); anti-phospho-STAT3 (Tyr705) (1:1000, #9131 from Cell Signaling); anti-GAPDH (1:1000, #2118 from Cell Signaling); anti-NOS2 (1:1000, ab178945 from Abcam); anti-IgG (1:1000, #7076 from Cell Signaling).

    Techniques: Western Blot, Pyrolysis Gas Chromatography, Mouse Assay

    ( A ) Representative Western blot of PGC-1α in the livers of PGC-1α +/+ (WT—wild-type) and PGC-1α -/- (KO—knockout) mice. Β-tubulin was used as the loading control. ( B ) mRNA relative expression of Tnfa , Il6 and Nos2 versus Tbp (TATA binding protein; housekeeping) in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced acute pancreatitis (AP) (Cerulein). ( C ) Representative Western blot of NOS2 in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). GAPDH was used as the loading control. ( D ) mRNA relative levels of Nos2 versus Tbp in the pancreas of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). ( E ) Representative Western blot of NOS2 in the pancreas of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). Β-tubulin was used as the loading control. There were six mice per group. Statistical difference is indicated as p *

    Journal: Antioxidants

    Article Title: Impairment of PGC-1 Alpha Up-Regulation Enhances Nitrosative Stress in the Liver during Acute Pancreatitis in Obese Mice

    doi: 10.3390/antiox9090887

    Figure Lengend Snippet: ( A ) Representative Western blot of PGC-1α in the livers of PGC-1α +/+ (WT—wild-type) and PGC-1α -/- (KO—knockout) mice. Β-tubulin was used as the loading control. ( B ) mRNA relative expression of Tnfa , Il6 and Nos2 versus Tbp (TATA binding protein; housekeeping) in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced acute pancreatitis (AP) (Cerulein). ( C ) Representative Western blot of NOS2 in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). GAPDH was used as the loading control. ( D ) mRNA relative levels of Nos2 versus Tbp in the pancreas of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). ( E ) Representative Western blot of NOS2 in the pancreas of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). Β-tubulin was used as the loading control. There were six mice per group. Statistical difference is indicated as p *

    Article Snippet: The employed antibodies were: anti-β-tubulin (1:1000, ab6046 from Abcam, Cambridge, UK); anti-PGC-1α (1:500, sc-518025 from Santa Cruz Biotechnology, Dallas, TX, USA); anti-p65 (1:1000, #8242 from Cell Signaling Technology, Danvers, MA, USA); anti-phospho-p65 (Ser 536) (1:100, #3033 from Cell Signaling); anti-Nitro-tyrosine (1:1000, #9691 from Cell Signaling); anti-STAT3 (1:1000, #9132 from Cell Signaling); anti-phospho-STAT3 (Tyr705) (1:1000, #9131 from Cell Signaling); anti-GAPDH (1:1000, #2118 from Cell Signaling); anti-NOS2 (1:1000, ab178945 from Abcam); anti-IgG (1:1000, #7076 from Cell Signaling).

    Techniques: Western Blot, Pyrolysis Gas Chromatography, Knock-Out, Mouse Assay, Expressing, Binding Assay

    ( A ) mRNA relative expression of Ppargc1a versus Tbp (TATA-binding protein; housekeeping) in the livers of the control (Sham) and 1 h after the cerulein-induced acute pancreatitis (Cerulein) mice. ( B ) Representative Western blot of PGC-1α in the livers of the control (Sham) and 1 h after the cerulein-induced acute pancreatitis (Cerulein) mice. β-tubulin was used as the loading control. There were six mice per group. Statistical difference is indicated as * p

    Journal: Antioxidants

    Article Title: Impairment of PGC-1 Alpha Up-Regulation Enhances Nitrosative Stress in the Liver during Acute Pancreatitis in Obese Mice

    doi: 10.3390/antiox9090887

    Figure Lengend Snippet: ( A ) mRNA relative expression of Ppargc1a versus Tbp (TATA-binding protein; housekeeping) in the livers of the control (Sham) and 1 h after the cerulein-induced acute pancreatitis (Cerulein) mice. ( B ) Representative Western blot of PGC-1α in the livers of the control (Sham) and 1 h after the cerulein-induced acute pancreatitis (Cerulein) mice. β-tubulin was used as the loading control. There were six mice per group. Statistical difference is indicated as * p

    Article Snippet: The employed antibodies were: anti-β-tubulin (1:1000, ab6046 from Abcam, Cambridge, UK); anti-PGC-1α (1:500, sc-518025 from Santa Cruz Biotechnology, Dallas, TX, USA); anti-p65 (1:1000, #8242 from Cell Signaling Technology, Danvers, MA, USA); anti-phospho-p65 (Ser 536) (1:100, #3033 from Cell Signaling); anti-Nitro-tyrosine (1:1000, #9691 from Cell Signaling); anti-STAT3 (1:1000, #9132 from Cell Signaling); anti-phospho-STAT3 (Tyr705) (1:1000, #9131 from Cell Signaling); anti-GAPDH (1:1000, #2118 from Cell Signaling); anti-NOS2 (1:1000, ab178945 from Abcam); anti-IgG (1:1000, #7076 from Cell Signaling).

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

    ( A ) Representative Western blot of 3-Nitrotyrosine (NT) in the pancreas and liver of the control (Sham) and 1 h after cerulein-induced acute pancreatitis (Cerulein) mice. Ponceau was used as the loading control. ( B ) Determination of the 3NO2-Tyr/p-Tyr ratio in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). ( C ) Representative Western blot of 3-NT in the liver after inducing acute pancreatitis (Cerulein) in the PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice. Ponceau was used as the loading control. ( D ) Energy charge in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). There were six mice per group. The statistical difference is indicated as ** p

    Journal: Antioxidants

    Article Title: Impairment of PGC-1 Alpha Up-Regulation Enhances Nitrosative Stress in the Liver during Acute Pancreatitis in Obese Mice

    doi: 10.3390/antiox9090887

    Figure Lengend Snippet: ( A ) Representative Western blot of 3-Nitrotyrosine (NT) in the pancreas and liver of the control (Sham) and 1 h after cerulein-induced acute pancreatitis (Cerulein) mice. Ponceau was used as the loading control. ( B ) Determination of the 3NO2-Tyr/p-Tyr ratio in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). ( C ) Representative Western blot of 3-NT in the liver after inducing acute pancreatitis (Cerulein) in the PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice. Ponceau was used as the loading control. ( D ) Energy charge in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). There were six mice per group. The statistical difference is indicated as ** p

    Article Snippet: The employed antibodies were: anti-β-tubulin (1:1000, ab6046 from Abcam, Cambridge, UK); anti-PGC-1α (1:500, sc-518025 from Santa Cruz Biotechnology, Dallas, TX, USA); anti-p65 (1:1000, #8242 from Cell Signaling Technology, Danvers, MA, USA); anti-phospho-p65 (Ser 536) (1:100, #3033 from Cell Signaling); anti-Nitro-tyrosine (1:1000, #9691 from Cell Signaling); anti-STAT3 (1:1000, #9132 from Cell Signaling); anti-phospho-STAT3 (Tyr705) (1:1000, #9131 from Cell Signaling); anti-GAPDH (1:1000, #2118 from Cell Signaling); anti-NOS2 (1:1000, ab178945 from Abcam); anti-IgG (1:1000, #7076 from Cell Signaling).

    Techniques: Western Blot, Mouse Assay, Pyrolysis Gas Chromatography

    ( A ) mRNA relative expression of Sod2 and Prx3 versus the Tbp (TATA-binding protein; housekeeping) in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). ( B ) The GSSG/GSH, g-glutamilcystine/g-glutamilcysteine and cystine/cysteine ratios in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). There were six mice per group. The statistical difference is indicated as * p

    Journal: Antioxidants

    Article Title: Impairment of PGC-1 Alpha Up-Regulation Enhances Nitrosative Stress in the Liver during Acute Pancreatitis in Obese Mice

    doi: 10.3390/antiox9090887

    Figure Lengend Snippet: ( A ) mRNA relative expression of Sod2 and Prx3 versus the Tbp (TATA-binding protein; housekeeping) in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). ( B ) The GSSG/GSH, g-glutamilcystine/g-glutamilcysteine and cystine/cysteine ratios in the livers of the sham PGC-1α +/+ (WT) and PGC-1α -/- (KO) mice and at 1 h after cerulein-induced AP (Cerulein). There were six mice per group. The statistical difference is indicated as * p

    Article Snippet: The employed antibodies were: anti-β-tubulin (1:1000, ab6046 from Abcam, Cambridge, UK); anti-PGC-1α (1:500, sc-518025 from Santa Cruz Biotechnology, Dallas, TX, USA); anti-p65 (1:1000, #8242 from Cell Signaling Technology, Danvers, MA, USA); anti-phospho-p65 (Ser 536) (1:100, #3033 from Cell Signaling); anti-Nitro-tyrosine (1:1000, #9691 from Cell Signaling); anti-STAT3 (1:1000, #9132 from Cell Signaling); anti-phospho-STAT3 (Tyr705) (1:1000, #9131 from Cell Signaling); anti-GAPDH (1:1000, #2118 from Cell Signaling); anti-NOS2 (1:1000, ab178945 from Abcam); anti-IgG (1:1000, #7076 from Cell Signaling).

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

    Proposed model whereby AngII infusion leads to mitochondrial dysfunction and skeletal muscle wasting. Parallel to the well-characterized AngII-mediated activation of the FOXO-E3-UPS axis via inhibition of Akt, AngII also induces expression of the phosphatase PP2Cα, which dephosphorylates and inactivates AMPK. This leads to reduced PGC-1α, NRF1, and TFAM expression (less mitochondrial biogenesis), and reduced ULK1 activity. The AngII-mediated reduction in ULK1 activation inhibits a critical early step in the autophagy pathway and prevents recycling of damaged mitochondria (mitophagy). AngII also inhibits both mitochondrial fission and fusion through predominately AMPK-independent pathways, which likely contribute to mitochondrial dysfunction caused by elevated AngII. Prolonged mitochondrial dysfunction and energy depletion ultimately leads to release of caspase-3, initiation of apoptosis, and wasting.

    Journal: Skeletal Muscle

    Article Title: Protein phosphatase 2C-alpha knockdown reduces angiotensin II-mediated skeletal muscle wasting via restoration of mitochondrial recycling and function

    doi: 10.1186/2044-5040-4-20

    Figure Lengend Snippet: Proposed model whereby AngII infusion leads to mitochondrial dysfunction and skeletal muscle wasting. Parallel to the well-characterized AngII-mediated activation of the FOXO-E3-UPS axis via inhibition of Akt, AngII also induces expression of the phosphatase PP2Cα, which dephosphorylates and inactivates AMPK. This leads to reduced PGC-1α, NRF1, and TFAM expression (less mitochondrial biogenesis), and reduced ULK1 activity. The AngII-mediated reduction in ULK1 activation inhibits a critical early step in the autophagy pathway and prevents recycling of damaged mitochondria (mitophagy). AngII also inhibits both mitochondrial fission and fusion through predominately AMPK-independent pathways, which likely contribute to mitochondrial dysfunction caused by elevated AngII. Prolonged mitochondrial dysfunction and energy depletion ultimately leads to release of caspase-3, initiation of apoptosis, and wasting.

    Article Snippet: AngII inhibited AMPK activity and reduced PGC-1α and TFAM expression (thereby inhibiting mitochondrial biogenesis) and impaired ULK1 activation and autophagy (thereby also inhibiting clearance of damaged mitochondria), resulting in mitochondrial dysfunction, decreased ATP, and wasting.

    Techniques: Activation Assay, Inhibition, Expressing, Pyrolysis Gas Chromatography, Activity Assay

    PP2Cα knockdown restored AMPK activation and signaling related to mitochondrial biogenesis. Specificity was ensured by performing experiments with two siRNA target sequences, as indicated. (A) Representative western blots showing effects of AngII and PP2Cα siRNA ‘A’ on the AMPK-PGC-1α-TFAM signaling axis. (B) PP2Cα protein expression was increased with AngII, which was prevented with PP2Cα siRNA. (C) Activating AMPK phosphorylation was reduced with AngII and increased with PP2Cα knockdown. (D) PGC-1α protein expression was reduced with AngII, which was prevented with PP2Cα siRNA. (E) PP2Cα siRNA increased expression of NRF1 transcription factor. (F) AngII reduced expression of the mitochondrial transcription factor TFAM, and this reduction was prevented by PP2Cα siRNA. n =4-32 per group, Mean ± SEM, * P

    Journal: Skeletal Muscle

    Article Title: Protein phosphatase 2C-alpha knockdown reduces angiotensin II-mediated skeletal muscle wasting via restoration of mitochondrial recycling and function

    doi: 10.1186/2044-5040-4-20

    Figure Lengend Snippet: PP2Cα knockdown restored AMPK activation and signaling related to mitochondrial biogenesis. Specificity was ensured by performing experiments with two siRNA target sequences, as indicated. (A) Representative western blots showing effects of AngII and PP2Cα siRNA ‘A’ on the AMPK-PGC-1α-TFAM signaling axis. (B) PP2Cα protein expression was increased with AngII, which was prevented with PP2Cα siRNA. (C) Activating AMPK phosphorylation was reduced with AngII and increased with PP2Cα knockdown. (D) PGC-1α protein expression was reduced with AngII, which was prevented with PP2Cα siRNA. (E) PP2Cα siRNA increased expression of NRF1 transcription factor. (F) AngII reduced expression of the mitochondrial transcription factor TFAM, and this reduction was prevented by PP2Cα siRNA. n =4-32 per group, Mean ± SEM, * P

    Article Snippet: AngII inhibited AMPK activity and reduced PGC-1α and TFAM expression (thereby inhibiting mitochondrial biogenesis) and impaired ULK1 activation and autophagy (thereby also inhibiting clearance of damaged mitochondria), resulting in mitochondrial dysfunction, decreased ATP, and wasting.

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