Western Blot:Article Title: Heart-Specific and Conditional Deletion of the Immt Gene Reveals Its Role in Regulating Mitochondrial Structure and Total Heart Metabolism
Article Snippet: The membranes were then washed with TBST and incubated with secondary indicator antibody (IRDye secondary antibodies, LI-COR, 1:3000) in 5% milk protein/TBST with 0.01% SDS at room temperature for 1 h. After washing with TBST 3 times, the membranes were then scanned by the Odyssey CLx imaging system with Image Studio software version 6.1 (LI-COR, Lincoln, NE, USA). .. Primary antibodies used for Western blots were anti-Mitofilin (Mic60 or Immt) (#10179-1-AP, Proteintech, Rosemont, IL, USA, 1:1000), anti-Mic19 (#25625-1-AP, Proteintech, 1:1000), anti-Sam50 (#28679-1-AP, Proteintech, 1:1000), anti-PINK1 (#BC100-494, Novus Biologicals, Centennial, CO, USA, 1:1000), anti-LC3 (#3868, Cell Signaling Technology, Danvers, MA, USA, 1:1000), anti-p62/SQSTM1 (#P0067, Sigma Aldrich, 1:1000), anti-LAMP2 (#ABL-93, Developmental Studies Hybridoma Bank, 1:50), anti-LIMPII (#ab176317, abcam, Cambridge, UK, 1:1000), anti-PGC1α (#ab54481, abcam, 1:1000), anti-PGC1β (#ab176328, abcam 1:1000), anti-MFN2 (#9482, Cell Signaling Technology, 1:1000), anti-MFN1 (#ab104274, abcam, 1:750), anti-Ubiquitin (#sc-8017, Santa Cruz, Dallas, TX, USA, 1:200), anti-GAPDH (#10R-G109A, Fitzgerald, Acton, MA, USA, 1:5000), anti-Mic26 (#MA5-15493, Thermo Fisher Scientific, Waltham, MA, USA, 1:1000), antiMic27 (#28514-1-AP, Proteintech, 1:1000), anti-Mic10 (#31561-1-AP, Proteintech, 1:500), anti-SESN2 (#10795-1-AP, Proteintech, 1:1000), anti-cGas (#31659, Cell Signaling Technology, 1:1000), anti-cMYC (#10828-1-AP, Proteintech, 1:2000), anti-GSDMA (#30354-1-AP, Proteintech, 1:2000), anti-OPTN (#10837-1-AP, Proteintech, 1:5000), anti-OPA1 (#ab157457, abcam, 1:1000), and anti-DRP1 (#8570, Cell Signaling Technology, 1:1000). .. Total RNA was isolated with TRIzol reagent (#15596018, Thermo Fisher Scientific) from heart tissue as previously described [25].
Article Title: Heart-Specific and Conditional Deletion of the Immt Gene Reveals Its Role in Regulating Mitochondrial Structure and Total Heart Metabolism
Article Snippet: The membranes were then washed with TBST and incubated with secondary indicator antibody (IRDye secondary antibodies, LI-COR, 1:3000) in 5% milk protein/TBST with 0.01% SDS at room temperature for 1 h. After washing with TBST 3 times, the membranes were then scanned by the Odyssey CLx imaging system with Image Studio software version 6.1 (LI-COR, Lincoln, NE, USA). .. Primary antibodies used for Western blots were anti-Mitofilin (Mic60 or Immt) (#10179-1-AP, Proteintech, Rosemont, IL, USA, 1:1000), anti-Mic19 (#25625-1-AP, Proteintech, 1:1000), anti-Sam50 (#28679-1-AP, Proteintech, 1:1000), anti-PINK1 (#BC100-494, Novus Biologicals, Centennial, CO, USA, 1:1000), anti-LC3 (#3868, Cell Signaling Technology, Danvers, MA, USA, 1:1000), anti-p62/SQSTM1 (#P0067, Sigma Aldrich, 1:1000), anti-LAMP2 (#ABL-93, Developmental Studies Hybridoma Bank, 1:50), anti-LIMPII (#ab176317, abcam, Cambridge, UK, 1:1000), anti-PGC1α (#ab54481, abcam, 1:1000), anti-PGC1β (#ab176328, abcam 1:1000), anti-MFN2 (#9482, Cell Signaling Technology, 1:1000), anti-MFN1 (#ab104274, abcam, 1:750), anti-Ubiquitin (#sc-8017, Santa Cruz, Dallas, TX, USA, 1:200), anti-GAPDH (#10R-G109A, Fitzgerald, Acton, MA, USA, 1:5000), anti-Mic26 (#MA5-15493, Thermo Fisher Scientific, Waltham, MA, USA, 1:1000), anti-Mic27 (#28514-1-AP, Proteintech, 1:1000), anti-Mic10 (#31561-1-AP, Proteintech, 1:500), anti-SESN2 (#10795-1-AP, Proteintech, 1:1000), anti-cGas (#31659, Cell Signaling Technology, 1:1000), anti-cMYC (#10828-1-AP, Proteintech, 1:2000), anti-GSDMA (#30354-1-AP, Proteintech, 1:2000), anti-OPTN (#10837-1-AP, Proteintech, 1:5000), anti-OPA1 (#ab157457, abcam, 1:1000), and anti-DRP1 (#8570, Cell Signaling Technology, 1:1000). .. Total RNA was isolated with TRIzol reagent (#15596018, Thermo Fisher Scientific) from heart tissue as previously described [ ].
Article Title: Cytosolic acetyl-coenzyme A is a signalling metabolite to control mitophagy.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Anti-TIM23 (mouse, 611223, BD Biosciences, 1:5000 dilution), anti-MT-CO2 (rabbit, ab79393, Abcam, 1:3000 dilution), anti-CYTB (rabbit, 55090-1-AP, Proteintech, 1:5000 dilution), anti- HSP60 (goat, sc-13115, Santa Cruz Biotechnologies, 1:1000 dilution), anti-LC3 (rabbit, 3868S, CST, 1:1000 dilution), anti-HA (mouse, 901513, Biolegend, 1:1000 dilution), anti-FLAG (mouse, F3165, Sigma, 1:3000), anti-NLRX1 (rabbit, 17215-1-AP, Proteintech, 1:1000 dilution), anti-PINK1 (rabbit, BC100-494SS, Novus Biologicals, 1:1000 dilution), anti-ACLY (rabbit, 15421-1-AP, Proteintech, 1:1000 dilution), anti-ACSS2 (rabbit, 16087-1-AP, Proteintech, 1:1000 dilution), anti-FASN (rabbit, 10624-2-AP, Proteintech, 1:3000 dilution), anti- ACC1 (rabbit, 21923-1-AP, Proteintech, 1:2000 dilution), anti-IDH1 (Rabbit, 12332-1-AP, proteintech, 1:3000 dilution), anti-p62 (rabbit, 18420-1-AP, Proteintech, 1:3000 dilution), anti-Acetylated-Lysine (rabbit, 9441, CST, 1:1000 dilution), anti-phospho-AMPKα (rabbit, 40H9, CST, 1:1000 dilution), anti-AMPKα (rabbit, 10929-2-AP, Proteintech, 1:3000 dilution), anti-ULK1 (Rabbit, #8054, CST, 1:1000 dilution), anti-phospho-ULK1 (Ser757) ( Rabbit, #6888, CST, 1:1000 dilution), anti-Phospho-ULK1 (Ser555) (D1H4) (Rabbit, #5869, CST, 1:1000 dilution), anti-S6 Kinase (S6K) (Rabbit, #9202, CST, 1:1000 dilution), anti-phospho-S6K (Thr389) (Mouse, #9206, CST,1:1000 dilution), anti-Cytochrome c (rabbit, 556432, BD Biosciences, 1:1000 dilution), anti-Parkin (rabbit, Proteintech, 66674-1-Ig, 1:500 dilution), anti-CLIMP63 (mouse, ENZ-ABS-669-0100, ENZO, 1:500 dilution), antiFIP200 (rabbit, 17250-1-AP, Proteintech, 1:3000 dilution), anti-ATG7 (rabbit, 10088-2-AP, Proteintech, 1:3000 dilution), anti-Tubulin (rabbit, 11224-1-AP, Proteintech, 1:5000 dilution), anti-ACTIN (mouse, 66009-1-Ig, Proteintech, 1:5000 dilution) were used in immunoblotting. .. Anti-LC3 (rabbit, PM036, MBL, 1:100 dilution), anti-HA (mouse, 901513, Biolegend, 1:1000 dilution), anti-TOM20 (mouse, 612278, BD Biosciences, 1:1000 dilution) and anti-HSP60 (goat, sc-13115, Santa Cruz Biotechnologies, 1:1000 dilution) were used in immunofluorescence.
other:
Incubation:Article Title: Mitochondrial transplantation ameliorates experimental autoimmune encephalomyelitis by modulating the Th17/Treg balance and restoring metabolic homeostasis
Article Snippet: .. The sections were then incubated overnight at 4 °C with the following primary antibodies at a 1:200 dilution: anti-TOM20 (ab56783; abcam), anti-TFAM (22586-1-AP; Proteintech), anti-PINK1 (BC100-494; Novus Biological), anti-IL-17 (ab79056; abcam), and anti-TNFα (ab6671; abcam), anti-iNOS (PA3-030A; Invitrogen), anti-COL1A1 (PA5-29569; Invitrogen), and anti-α-SMA (ab7817; abcam), Sections were incubated with secondary biotinylated antibodies, then incubated with streptavidin–peroxidase complex for 30 min. .. Reaction products were developed using 3,3-diaminobenzidine chromogen (K3468; Dako Corp.).
Article Title: Kisspeptin-10 attenuates pulmonary arterial hypertension via restoration of mitochondrial function in pulmonary artery smooth muscle cells
Article Snippet: Mitochondrial dysfunction and dysregulated mitophagy in pulmonary artery smooth muscle cells (PASMCs) critically drive pulmonary arterial hypertension (PAH).. Here we show that Kisspeptin-10, acting via its receptor GPR54, markedly attenuates PAH in the SU5416/hypoxia mouse model.. The administration of kisspeptin-10 resulted in a considerable reduction in the systolic pressure of the right ventricle, as well as in the level of right ventricular hypertrophy and pulmonary vascular remodeling.
Chromatin Immunoprecipitation:Article Title: Cytosolic acetyl-coenzyme A is a signalling metabolite to control mitophagy.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Anti-TIM23 (mouse, 611223, BD Biosciences, 1:5000 dilution), anti-MT-CO2 (rabbit, ab79393, Abcam, 1:3000 dilution), anti-CYTB (rabbit, 55090-1-AP, Proteintech, 1:5000 dilution), anti- HSP60 (goat, sc-13115, Santa Cruz Biotechnologies, 1:1000 dilution), anti-LC3 (rabbit, 3868S, CST, 1:1000 dilution), anti-HA (mouse, 901513, Biolegend, 1:1000 dilution), anti-FLAG (mouse, F3165, Sigma, 1:3000), anti-NLRX1 (rabbit, 17215-1-AP, Proteintech, 1:1000 dilution), anti-PINK1 (rabbit, BC100-494SS, Novus Biologicals, 1:1000 dilution), anti-ACLY (rabbit, 15421-1-AP, Proteintech, 1:1000 dilution), anti-ACSS2 (rabbit, 16087-1-AP, Proteintech, 1:1000 dilution), anti-FASN (rabbit, 10624-2-AP, Proteintech, 1:3000 dilution), anti- ACC1 (rabbit, 21923-1-AP, Proteintech, 1:2000 dilution), anti-IDH1 (Rabbit, 12332-1-AP, proteintech, 1:3000 dilution), anti-p62 (rabbit, 18420-1-AP, Proteintech, 1:3000 dilution), anti-Acetylated-Lysine (rabbit, 9441, CST, 1:1000 dilution), anti-phospho-AMPKα (rabbit, 40H9, CST, 1:1000 dilution), anti-AMPKα (rabbit, 10929-2-AP, Proteintech, 1:3000 dilution), anti-ULK1 (Rabbit, #8054, CST, 1:1000 dilution), anti-phospho-ULK1 (Ser757) ( Rabbit, #6888, CST, 1:1000 dilution), anti-Phospho-ULK1 (Ser555) (D1H4) (Rabbit, #5869, CST, 1:1000 dilution), anti-S6 Kinase (S6K) (Rabbit, #9202, CST, 1:1000 dilution), anti-phospho-S6K (Thr389) (Mouse, #9206, CST,1:1000 dilution), anti-Cytochrome c (rabbit, 556432, BD Biosciences, 1:1000 dilution), anti-Parkin (rabbit, Proteintech, 66674-1-Ig, 1:500 dilution), anti-CLIMP63 (mouse, ENZ-ABS-669-0100, ENZO, 1:500 dilution), antiFIP200 (rabbit, 17250-1-AP, Proteintech, 1:3000 dilution), anti-ATG7 (rabbit, 10088-2-AP, Proteintech, 1:3000 dilution), anti-Tubulin (rabbit, 11224-1-AP, Proteintech, 1:5000 dilution), anti-ACTIN (mouse, 66009-1-Ig, Proteintech, 1:5000 dilution) were used in immunoblotting. .. Anti-LC3 (rabbit, PM036, MBL, 1:100 dilution), anti-HA (mouse, 901513, Biolegend, 1:1000 dilution), anti-TOM20 (mouse, 612278, BD Biosciences, 1:1000 dilution) and anti-HSP60 (goat, sc-13115, Santa Cruz Biotechnologies, 1:1000 dilution) were used in immunofluorescence.
Flow Cytometry:Article Title: Cytosolic acetyl-coenzyme A is a signalling metabolite to control mitophagy.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Anti-TIM23 (mouse, 611223, BD Biosciences, 1:5000 dilution), anti-MT-CO2 (rabbit, ab79393, Abcam, 1:3000 dilution), anti-CYTB (rabbit, 55090-1-AP, Proteintech, 1:5000 dilution), anti- HSP60 (goat, sc-13115, Santa Cruz Biotechnologies, 1:1000 dilution), anti-LC3 (rabbit, 3868S, CST, 1:1000 dilution), anti-HA (mouse, 901513, Biolegend, 1:1000 dilution), anti-FLAG (mouse, F3165, Sigma, 1:3000), anti-NLRX1 (rabbit, 17215-1-AP, Proteintech, 1:1000 dilution), anti-PINK1 (rabbit, BC100-494SS, Novus Biologicals, 1:1000 dilution), anti-ACLY (rabbit, 15421-1-AP, Proteintech, 1:1000 dilution), anti-ACSS2 (rabbit, 16087-1-AP, Proteintech, 1:1000 dilution), anti-FASN (rabbit, 10624-2-AP, Proteintech, 1:3000 dilution), anti- ACC1 (rabbit, 21923-1-AP, Proteintech, 1:2000 dilution), anti-IDH1 (Rabbit, 12332-1-AP, proteintech, 1:3000 dilution), anti-p62 (rabbit, 18420-1-AP, Proteintech, 1:3000 dilution), anti-Acetylated-Lysine (rabbit, 9441, CST, 1:1000 dilution), anti-phospho-AMPKα (rabbit, 40H9, CST, 1:1000 dilution), anti-AMPKα (rabbit, 10929-2-AP, Proteintech, 1:3000 dilution), anti-ULK1 (Rabbit, #8054, CST, 1:1000 dilution), anti-phospho-ULK1 (Ser757) ( Rabbit, #6888, CST, 1:1000 dilution), anti-Phospho-ULK1 (Ser555) (D1H4) (Rabbit, #5869, CST, 1:1000 dilution), anti-S6 Kinase (S6K) (Rabbit, #9202, CST, 1:1000 dilution), anti-phospho-S6K (Thr389) (Mouse, #9206, CST,1:1000 dilution), anti-Cytochrome c (rabbit, 556432, BD Biosciences, 1:1000 dilution), anti-Parkin (rabbit, Proteintech, 66674-1-Ig, 1:500 dilution), anti-CLIMP63 (mouse, ENZ-ABS-669-0100, ENZO, 1:500 dilution), antiFIP200 (rabbit, 17250-1-AP, Proteintech, 1:3000 dilution), anti-ATG7 (rabbit, 10088-2-AP, Proteintech, 1:3000 dilution), anti-Tubulin (rabbit, 11224-1-AP, Proteintech, 1:5000 dilution), anti-ACTIN (mouse, 66009-1-Ig, Proteintech, 1:5000 dilution) were used in immunoblotting. .. Anti-LC3 (rabbit, PM036, MBL, 1:100 dilution), anti-HA (mouse, 901513, Biolegend, 1:1000 dilution), anti-TOM20 (mouse, 612278, BD Biosciences, 1:1000 dilution) and anti-HSP60 (goat, sc-13115, Santa Cruz Biotechnologies, 1:1000 dilution) were used in immunofluorescence.
Magnetic Resonance Imaging:Article Title: Cytosolic acetyl-coenzyme A is a signalling metabolite to control mitophagy.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Anti-TIM23 (mouse, 611223, BD Biosciences, 1:5000 dilution), anti-MT-CO2 (rabbit, ab79393, Abcam, 1:3000 dilution), anti-CYTB (rabbit, 55090-1-AP, Proteintech, 1:5000 dilution), anti- HSP60 (goat, sc-13115, Santa Cruz Biotechnologies, 1:1000 dilution), anti-LC3 (rabbit, 3868S, CST, 1:1000 dilution), anti-HA (mouse, 901513, Biolegend, 1:1000 dilution), anti-FLAG (mouse, F3165, Sigma, 1:3000), anti-NLRX1 (rabbit, 17215-1-AP, Proteintech, 1:1000 dilution), anti-PINK1 (rabbit, BC100-494SS, Novus Biologicals, 1:1000 dilution), anti-ACLY (rabbit, 15421-1-AP, Proteintech, 1:1000 dilution), anti-ACSS2 (rabbit, 16087-1-AP, Proteintech, 1:1000 dilution), anti-FASN (rabbit, 10624-2-AP, Proteintech, 1:3000 dilution), anti- ACC1 (rabbit, 21923-1-AP, Proteintech, 1:2000 dilution), anti-IDH1 (Rabbit, 12332-1-AP, proteintech, 1:3000 dilution), anti-p62 (rabbit, 18420-1-AP, Proteintech, 1:3000 dilution), anti-Acetylated-Lysine (rabbit, 9441, CST, 1:1000 dilution), anti-phospho-AMPKα (rabbit, 40H9, CST, 1:1000 dilution), anti-AMPKα (rabbit, 10929-2-AP, Proteintech, 1:3000 dilution), anti-ULK1 (Rabbit, #8054, CST, 1:1000 dilution), anti-phospho-ULK1 (Ser757) ( Rabbit, #6888, CST, 1:1000 dilution), anti-Phospho-ULK1 (Ser555) (D1H4) (Rabbit, #5869, CST, 1:1000 dilution), anti-S6 Kinase (S6K) (Rabbit, #9202, CST, 1:1000 dilution), anti-phospho-S6K (Thr389) (Mouse, #9206, CST,1:1000 dilution), anti-Cytochrome c (rabbit, 556432, BD Biosciences, 1:1000 dilution), anti-Parkin (rabbit, Proteintech, 66674-1-Ig, 1:500 dilution), anti-CLIMP63 (mouse, ENZ-ABS-669-0100, ENZO, 1:500 dilution), antiFIP200 (rabbit, 17250-1-AP, Proteintech, 1:3000 dilution), anti-ATG7 (rabbit, 10088-2-AP, Proteintech, 1:3000 dilution), anti-Tubulin (rabbit, 11224-1-AP, Proteintech, 1:5000 dilution), anti-ACTIN (mouse, 66009-1-Ig, Proteintech, 1:5000 dilution) were used in immunoblotting. .. Anti-LC3 (rabbit, PM036, MBL, 1:100 dilution), anti-HA (mouse, 901513, Biolegend, 1:1000 dilution), anti-TOM20 (mouse, 612278, BD Biosciences, 1:1000 dilution) and anti-HSP60 (goat, sc-13115, Santa Cruz Biotechnologies, 1:1000 dilution) were used in immunofluorescence.
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