Knockdown:Article Title: Chchd10: A Novel Metabolic Sensor Modulating Adipose Tissue Homeostasis
Article Snippet: .. [ ] Briefly, IR was induced with the treatment of both TNFα (2.5 n m , 410‐MT, R&D System) and hypoxia (1% oxygen) for 16 h. To knock down GSTA4 expression, 2‐day‐differentiated Chchd10 KO and WT adipocytes were infected with adenovirus encoding GSTA4 shRNA (Ad‐shGSTA4) or scramble RNA (Ad‐scramble) for 48 h. To inhibit NRF2, p62, or mTORC1, 2‐day‐differentiated adipocytes were treated with ML385 (10μ m , HY‐100523, MedChemExpress), K67 (10μ m , HY‐111126, MedChemExpress), or rapamycin (200 nM, 553 210, Sigma‐Aldrich) for 48 h, respectively. .. To knock down NRF2 and TDP43 expression, 2‐day‐differentiated adipocytes were transfected with mouse NFE2L2 siRNA (siNRF2) targeting 3′ UTR (mm.Ri.Nfe2l2.13.1, Integrated DNA Technologies), mouse Tardbp siRNA (siTDP43) targeting exon 3 and 4 (mm.Ri.Tardbp.13.2, Integrated DNA Technologies), or scramble siRNA as control (siCTR, #51‐01‐14, Integrated DNA Technologies) using lipofectamine 3000 transfection system (Invitrogen) for 48 h. To assess Raptor RNA stability, 4‐day‐differentiated adipocytes were treated with actinomycin D (1μg mL −1 , A9415, Sigma‐Aldrich) or PBS for 0, 2, or 4 h. The RNA was then extracted, reverse transcribed into cDNA, and subjected to qPCR analysis.
Article Title: Chchd10: A Novel Metabolic Sensor Modulating Adipose Tissue Homeostasis.
Article Snippet: .. [42] Briefly, IR was induced with the treatment of both TNFα (2.5 nm, 410-MT, R&D System) and hypoxia (1% oxygen) for 16 h. To knock down GSTA4 expression, 2-day-differentiated Chchd10 KO and WT adipocytes were infected with adenovirus encoding GSTA4 shRNA (Ad-shGSTA4) or scramble RNA (Ad-scramble) for 48 h. To inhibit NRF2, p62, or mTORC1, 2- day-differentiated adipocytes were treated with ML385 (10μm, HY-100523, MedChemExpress), K67 (10μm, HY-111126, MedChemExpress), or rapamycin (200 nM, 553 210, Sigma-Aldrich) for 48 h, respectively. .. To knock down NRF2 and TDP43 expression, 2-day-differentiated adipocytes were transfected with mouse NFE2L2 siRNA (siNRF2) targeting 3′ UTR (mm.Ri.Nfe2l2.13.1, Integrated DNA Technologies), mouse Tardbp siRNA (siTDP43) targeting exon 3 and 4 (mm.Ri.Tardbp.13.2, Integrated DNA Technologies), or scramble siRNA as control (siCTR, #51-01-14, Integrated DNA Technologies) using lipofectamine 3000 transfection system (Invitrogen) for 48 h. To assess Raptor RNA stability, 4-day-differentiated adipocytes were treated with actinomycin D (1μg mL−1, A9415, SigmaAldrich) or PBS for 0, 2, or 4 h. The RNA was then extracted, reverse transcribed into cDNA, and subjected to qPCR analysis.
Expressing:Article Title: Chchd10: A Novel Metabolic Sensor Modulating Adipose Tissue Homeostasis
Article Snippet: .. [ ] Briefly, IR was induced with the treatment of both TNFα (2.5 n m , 410‐MT, R&D System) and hypoxia (1% oxygen) for 16 h. To knock down GSTA4 expression, 2‐day‐differentiated Chchd10 KO and WT adipocytes were infected with adenovirus encoding GSTA4 shRNA (Ad‐shGSTA4) or scramble RNA (Ad‐scramble) for 48 h. To inhibit NRF2, p62, or mTORC1, 2‐day‐differentiated adipocytes were treated with ML385 (10μ m , HY‐100523, MedChemExpress), K67 (10μ m , HY‐111126, MedChemExpress), or rapamycin (200 nM, 553 210, Sigma‐Aldrich) for 48 h, respectively. .. To knock down NRF2 and TDP43 expression, 2‐day‐differentiated adipocytes were transfected with mouse NFE2L2 siRNA (siNRF2) targeting 3′ UTR (mm.Ri.Nfe2l2.13.1, Integrated DNA Technologies), mouse Tardbp siRNA (siTDP43) targeting exon 3 and 4 (mm.Ri.Tardbp.13.2, Integrated DNA Technologies), or scramble siRNA as control (siCTR, #51‐01‐14, Integrated DNA Technologies) using lipofectamine 3000 transfection system (Invitrogen) for 48 h. To assess Raptor RNA stability, 4‐day‐differentiated adipocytes were treated with actinomycin D (1μg mL −1 , A9415, Sigma‐Aldrich) or PBS for 0, 2, or 4 h. The RNA was then extracted, reverse transcribed into cDNA, and subjected to qPCR analysis.
Article Title: Chchd10: A Novel Metabolic Sensor Modulating Adipose Tissue Homeostasis.
Article Snippet: .. [42] Briefly, IR was induced with the treatment of both TNFα (2.5 nm, 410-MT, R&D System) and hypoxia (1% oxygen) for 16 h. To knock down GSTA4 expression, 2-day-differentiated Chchd10 KO and WT adipocytes were infected with adenovirus encoding GSTA4 shRNA (Ad-shGSTA4) or scramble RNA (Ad-scramble) for 48 h. To inhibit NRF2, p62, or mTORC1, 2- day-differentiated adipocytes were treated with ML385 (10μm, HY-100523, MedChemExpress), K67 (10μm, HY-111126, MedChemExpress), or rapamycin (200 nM, 553 210, Sigma-Aldrich) for 48 h, respectively. .. To knock down NRF2 and TDP43 expression, 2-day-differentiated adipocytes were transfected with mouse NFE2L2 siRNA (siNRF2) targeting 3′ UTR (mm.Ri.Nfe2l2.13.1, Integrated DNA Technologies), mouse Tardbp siRNA (siTDP43) targeting exon 3 and 4 (mm.Ri.Tardbp.13.2, Integrated DNA Technologies), or scramble siRNA as control (siCTR, #51-01-14, Integrated DNA Technologies) using lipofectamine 3000 transfection system (Invitrogen) for 48 h. To assess Raptor RNA stability, 4-day-differentiated adipocytes were treated with actinomycin D (1μg mL−1, A9415, SigmaAldrich) or PBS for 0, 2, or 4 h. The RNA was then extracted, reverse transcribed into cDNA, and subjected to qPCR analysis.
Infection:Article Title: Chchd10: A Novel Metabolic Sensor Modulating Adipose Tissue Homeostasis
Article Snippet: .. [ ] Briefly, IR was induced with the treatment of both TNFα (2.5 n m , 410‐MT, R&D System) and hypoxia (1% oxygen) for 16 h. To knock down GSTA4 expression, 2‐day‐differentiated Chchd10 KO and WT adipocytes were infected with adenovirus encoding GSTA4 shRNA (Ad‐shGSTA4) or scramble RNA (Ad‐scramble) for 48 h. To inhibit NRF2, p62, or mTORC1, 2‐day‐differentiated adipocytes were treated with ML385 (10μ m , HY‐100523, MedChemExpress), K67 (10μ m , HY‐111126, MedChemExpress), or rapamycin (200 nM, 553 210, Sigma‐Aldrich) for 48 h, respectively. .. To knock down NRF2 and TDP43 expression, 2‐day‐differentiated adipocytes were transfected with mouse NFE2L2 siRNA (siNRF2) targeting 3′ UTR (mm.Ri.Nfe2l2.13.1, Integrated DNA Technologies), mouse Tardbp siRNA (siTDP43) targeting exon 3 and 4 (mm.Ri.Tardbp.13.2, Integrated DNA Technologies), or scramble siRNA as control (siCTR, #51‐01‐14, Integrated DNA Technologies) using lipofectamine 3000 transfection system (Invitrogen) for 48 h. To assess Raptor RNA stability, 4‐day‐differentiated adipocytes were treated with actinomycin D (1μg mL −1 , A9415, Sigma‐Aldrich) or PBS for 0, 2, or 4 h. The RNA was then extracted, reverse transcribed into cDNA, and subjected to qPCR analysis.
Article Title: Chchd10: A Novel Metabolic Sensor Modulating Adipose Tissue Homeostasis.
Article Snippet: .. [42] Briefly, IR was induced with the treatment of both TNFα (2.5 nm, 410-MT, R&D System) and hypoxia (1% oxygen) for 16 h. To knock down GSTA4 expression, 2-day-differentiated Chchd10 KO and WT adipocytes were infected with adenovirus encoding GSTA4 shRNA (Ad-shGSTA4) or scramble RNA (Ad-scramble) for 48 h. To inhibit NRF2, p62, or mTORC1, 2- day-differentiated adipocytes were treated with ML385 (10μm, HY-100523, MedChemExpress), K67 (10μm, HY-111126, MedChemExpress), or rapamycin (200 nM, 553 210, Sigma-Aldrich) for 48 h, respectively. .. To knock down NRF2 and TDP43 expression, 2-day-differentiated adipocytes were transfected with mouse NFE2L2 siRNA (siNRF2) targeting 3′ UTR (mm.Ri.Nfe2l2.13.1, Integrated DNA Technologies), mouse Tardbp siRNA (siTDP43) targeting exon 3 and 4 (mm.Ri.Tardbp.13.2, Integrated DNA Technologies), or scramble siRNA as control (siCTR, #51-01-14, Integrated DNA Technologies) using lipofectamine 3000 transfection system (Invitrogen) for 48 h. To assess Raptor RNA stability, 4-day-differentiated adipocytes were treated with actinomycin D (1μg mL−1, A9415, SigmaAldrich) or PBS for 0, 2, or 4 h. The RNA was then extracted, reverse transcribed into cDNA, and subjected to qPCR analysis.
shRNA:Article Title: Chchd10: A Novel Metabolic Sensor Modulating Adipose Tissue Homeostasis
Article Snippet: .. [ ] Briefly, IR was induced with the treatment of both TNFα (2.5 n m , 410‐MT, R&D System) and hypoxia (1% oxygen) for 16 h. To knock down GSTA4 expression, 2‐day‐differentiated Chchd10 KO and WT adipocytes were infected with adenovirus encoding GSTA4 shRNA (Ad‐shGSTA4) or scramble RNA (Ad‐scramble) for 48 h. To inhibit NRF2, p62, or mTORC1, 2‐day‐differentiated adipocytes were treated with ML385 (10μ m , HY‐100523, MedChemExpress), K67 (10μ m , HY‐111126, MedChemExpress), or rapamycin (200 nM, 553 210, Sigma‐Aldrich) for 48 h, respectively. .. To knock down NRF2 and TDP43 expression, 2‐day‐differentiated adipocytes were transfected with mouse NFE2L2 siRNA (siNRF2) targeting 3′ UTR (mm.Ri.Nfe2l2.13.1, Integrated DNA Technologies), mouse Tardbp siRNA (siTDP43) targeting exon 3 and 4 (mm.Ri.Tardbp.13.2, Integrated DNA Technologies), or scramble siRNA as control (siCTR, #51‐01‐14, Integrated DNA Technologies) using lipofectamine 3000 transfection system (Invitrogen) for 48 h. To assess Raptor RNA stability, 4‐day‐differentiated adipocytes were treated with actinomycin D (1μg mL −1 , A9415, Sigma‐Aldrich) or PBS for 0, 2, or 4 h. The RNA was then extracted, reverse transcribed into cDNA, and subjected to qPCR analysis.
Article Title: Chchd10: A Novel Metabolic Sensor Modulating Adipose Tissue Homeostasis.
Article Snippet: .. [42] Briefly, IR was induced with the treatment of both TNFα (2.5 nm, 410-MT, R&D System) and hypoxia (1% oxygen) for 16 h. To knock down GSTA4 expression, 2-day-differentiated Chchd10 KO and WT adipocytes were infected with adenovirus encoding GSTA4 shRNA (Ad-shGSTA4) or scramble RNA (Ad-scramble) for 48 h. To inhibit NRF2, p62, or mTORC1, 2- day-differentiated adipocytes were treated with ML385 (10μm, HY-100523, MedChemExpress), K67 (10μm, HY-111126, MedChemExpress), or rapamycin (200 nM, 553 210, Sigma-Aldrich) for 48 h, respectively. .. To knock down NRF2 and TDP43 expression, 2-day-differentiated adipocytes were transfected with mouse NFE2L2 siRNA (siNRF2) targeting 3′ UTR (mm.Ri.Nfe2l2.13.1, Integrated DNA Technologies), mouse Tardbp siRNA (siTDP43) targeting exon 3 and 4 (mm.Ri.Tardbp.13.2, Integrated DNA Technologies), or scramble siRNA as control (siCTR, #51-01-14, Integrated DNA Technologies) using lipofectamine 3000 transfection system (Invitrogen) for 48 h. To assess Raptor RNA stability, 4-day-differentiated adipocytes were treated with actinomycin D (1μg mL−1, A9415, SigmaAldrich) or PBS for 0, 2, or 4 h. The RNA was then extracted, reverse transcribed into cDNA, and subjected to qPCR analysis.
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