Membrane:Article Title: HIPK2 Confers Protection Against Septic Myocardial Injury by Regulating Ferroptosis.
Article Snippet: Sepsis causes systemic organ and tissue dysfunction, among which myocardial injury has become an urgent clinical problem.. Homeodomain-interacting protein kinase 2 (HIPK2) is a serine-threonine kinase that plays an important role in cellular activities.. In this study, we investigated the regulatory role of HIPK2 regarding the level of ferroptosis in septic myocardial injury.
Incubation:Article Title: HIPK2 Confers Protection Against Septic Myocardial Injury by Regulating Ferroptosis.
Article Snippet: Sepsis causes systemic organ and tissue dysfunction, among which myocardial injury has become an urgent clinical problem.. Homeodomain-interacting protein kinase 2 (HIPK2) is a serine-threonine kinase that plays an important role in cellular activities.. In this study, we investigated the regulatory role of HIPK2 regarding the level of ferroptosis in septic myocardial injury.
Pyrolysis Gas Chromatography:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Staining:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Immunohistochemical staining:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Multiple Displacement Amplification:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Enzyme-linked Immunosorbent Assay:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Isolation:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
CRISPR:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Activation Assay:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Plasmid Preparation:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Luciferase:Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis
Article Snippet: .. Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4-alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve-1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzyme-linked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C-11 BODIPY 581/591, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6 J (6–8 weeks old), Ppargc1α f/f (B6.Cg-Ppargc1α tm2.1Brsp /J) and transgenic (Tg) (Alb-cre) 21Mgn /J (Alb-cre +/+ ) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis.
Article Snippet: .. Materials and Methods Special reagents Special reagents included primary antibodies against Drp1, Nrf1, P-Drp1ser616, alpha-smooth muscle actin (α-SMA), collagen I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytochrome c oxidase subunit 4 (COXIV) and cleaved caspase-3 (Cell Signaling Technology, Beverly, USA); PGC-1α, ACSL4, tumor necrosis factor (TNF)α, Hepatocyte nuclear factor 4- alpha (HNF-4α), Desmin and interleukin (IL)-6 (Abcam, Cambridge, USA); Tim23 (Santa, TX, USA); glutathione peroxidase 4 (GPX4), Ferroton heavy chain 1 (FTH1), transferrin receptor 1 (TFR1) and F4/80 (Proteintech, Wuhan, China); Lymphatic Vessel Endothelial Receptor-1(Lyve1) (ABclonal, Wuhan, China); Nrf2, P-MLKL, GSDMD-N (HUABIO, Hangzhou, China); special kits for hematoxylin and eosin (H&E), Sirius Red, and Oil Red staining (Solarbio, Beijing, China); immunohistochemical staining kit (Maixin Biological Technology, Fujian, China); the kits for measurements of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), glucose, glutathione (GSH), malondialdehyde (MDA), and iron contents (Jiancheng Biological Engineering Institute, Nanjing, China); enzymelinked immunosorbent assay (ELISA) kits for measurements of insulin (Crystal Chem, Chicago, USA); Tissue mitochondria isolation kit (Beyotime, Shanghai, China); Perls’ blue (Sbjbio, Nanjing, China); PGC-1α clustered regularly interspaced short palindromic repeats (CRISPR) activation AR TI CL E IN P RE SS plasmid (sc-400070-ACT) and Protein A/G plus-agarose (Santa Cruz Biotechnology, CA, USA); PGC-1α and Drp1 small interfering RNAs (siRNAs) (LIKELI, Beijing, China); dual luciferase assay kits and TnT® quick coupled transcription/ translation systems (Promega, Wisconsin, USA); Lipofectamine 2000, C11 BODIPY, MitoSOX and Mitotracker (Invitrogen, CA, USA); JC-1 (Chemodex, SG, SUI ); Ferrostatin-1 (Fer-1) (MedChemExpress, NJ, USA). .. Male C57BL/6J (6–8 weeks old), Ppargc1αf/f (B6.Cg-Ppargc1αtm2.1Brsp/J) and transgenic (Tg) (Albcre)21Mgn/J (Alb-cre+/+) mice were from Vital River Laboratory Animal Technology (Beijing, China) and the Jackson Laboratory (Bar Harbor, ME, USA), respectively.
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