Journal: Cell Death & Disease

Article Title: Defective mitochondrial protein import contributes to complex I-induced mitochondrial dysfunction and neurodegeneration in Parkinson’s disease

doi: 10.1038/s41419-018-1154-0

Figure Lengend Snippet: a Representative immunoblots of TIM23 and TOM20 protein levels in substantia nigra homogenates from Ctrl ( n = 4–5) and PD patients ( n = 9). Protein levels were normalized relative to Ponceau-S. Quantification is depicted as fold change to Ctrl (* P < 0.05 after unpaired two-sided Student’s t-test). b ) Representative immunoblots of NDUFS3, COX IV and α-tubulin protein levels in substantia nigra homogenates from control (Ctrl, n = 5) and PD patients ( n = 9). Protein levels were normalized relative to α-tubulin (* P < 0.05 after unpaired two-sided Student’s t -test). Error bars indicate s.e.m.

Article Snippet: Human TIM23 and TOM20 plasmids cloned into a pCMV6-XL5 vector were obtained from Origene (Origene Technologies), respectively.

Techniques: Western Blot, Control

Journal: Cell Death & Disease

Article Title: Defective mitochondrial protein import contributes to complex I-induced mitochondrial dysfunction and neurodegeneration in Parkinson’s disease

doi: 10.1038/s41419-018-1154-0

Figure Lengend Snippet: a–c , e BE(2)-M17 cells were treated with MPP + (0.25, 0.5 or 1 mM as stated) or untreated (UT) for 24 h. a Representative immunoblots of TIM23, TOM20 and VDAC protein levels normalized relative to VDAC ( n = 3 independent experiments). Quantification is depicted as fold change to UT condition (* P < 0.05, ** P < 0.01 after one-way ANOVA test followed by Tukey’s post hoc test). b Representative images of mitoGFP-transfected BE(2)-M17 cells. Quantification is depicted as the fold change of mitoGFP intensity per cell compared with UT condition. A minimum of 35 cells were analyzed per condition (* P < 0.05 after unpaired two-sided Student’s t -test). Scale bar: 10 μm. c Representative immunoblots of GFP and β-actin protein levels. mitoGFP import was calculated as the ratio of mature (m) GFP compared to the sum of precursor (p) and mature GFP (total GFP; n = 5 independent experiments, *** P < 0.001 after unpaired two-sided Student’s t -test). d Representative immunoblots of radiolabelled OTC in isolated mitochondria UT or treated with MPP + (1 mM) at different time points. pOTC import was calculated as the percentage of mature (m) OTC compared to the sum of precursor (p) and mature OTC ( n = 3 independent experiments, *** P < 0.001 after unpaired two-sided Mann–Whitney test). e Representative images of NDUFS3 and COX IV immunofluorescence. Quantification is represented as the fold change in NDUFS3 or COX IV intensity compared with UT condition. A minimum of 35 cells were analyzed per condition (* P < 0.05, ** P < 0.01 after unpaired two-sided Student’s t -test or unpaired two-sided Mann–Whitney test). Scale bar: 10 μm. Error bars indicate s.e.m.

Article Snippet: Human TIM23 and TOM20 plasmids cloned into a pCMV6-XL5 vector were obtained from Origene (Origene Technologies), respectively.

Techniques: Western Blot, Transfection, Isolation, MANN-WHITNEY, Immunofluorescence

Journal: Cell Death & Disease

Article Title: Defective mitochondrial protein import contributes to complex I-induced mitochondrial dysfunction and neurodegeneration in Parkinson’s disease

doi: 10.1038/s41419-018-1154-0

Figure Lengend Snippet: Vehicle- or pCMV6-XL5-TIM23-transfected (TIM23) BE(2)-M17 cells were untreated (UT) or treated with a – c 1 mM MPP + for 24 h or d 5 mM for 24 h. a Representative images of COX IV and TIM23 immunofluorescence. Quantification is depicted as the fold change in COX IV intensity compared with vehicle-transfected UT condition. Dashed line shows TIM23-overexpressing cells. In all, 15 to 60 cells were analyzed per condition (* P < 0.05, *** P < 0.001 after two-way ANOVA followed by Tukey’s post hoc test). Scale bar: 10 μm. b ROS production measured as CM-H 2 DCFDA fluorescence intensity. Quantification is depicted as the fold change in fluorescence intensity compared with vehicle-transfected UT condition ( n = 3 independent experiments, ** P < 0.01 after one-way ANOVA followed by Tukey’s post hoc test). c Mitochondrial membrane potential measured as TMRM fluorescence intensity. Quantification is depicted as the fold change in fluorescence intensity compared with vehicle-transfected UT condition ( n = 5 independent experiments, ** P < 0.01 after one-way ANOVA followed by Tukey’s post hoc test). d Cell death measured as propidium iodide (PI)-positive cells ( n = 5 independent experiments). Quantification is depicted as the fold change in the percentage of PI-positive cells compared with each control condition (** P < 0.01 after unpaired two-sided Student’s t -test). Error bars indicate s.e.m.

Article Snippet: Human TIM23 and TOM20 plasmids cloned into a pCMV6-XL5 vector were obtained from Origene (Origene Technologies), respectively.

Techniques: Transfection, Immunofluorescence, Fluorescence, Membrane, Control

Journal: Cell Death & Disease

Article Title: Defective mitochondrial protein import contributes to complex I-induced mitochondrial dysfunction and neurodegeneration in Parkinson’s disease

doi: 10.1038/s41419-018-1154-0

Figure Lengend Snippet: a Representative immunoblots of TIM23, TOM20 and VDAC protein levels in ventral midbrain of vehicle- (Veh, n = 5) and MPTP-treated ( n = 6–7) mice euthanized at different time points. Protein levels were normalized relative to VDAC and quantification is depicted as fold change to vehicle condition (* P < 0.05, *** P < 0.001 after one-way ANOVA followed by Tukey’s post hoc test). b Representative images of tyrosine hydroxylase (TH) and COX IV immunostained in ventral midbrain sections from vehicle- ( n = 5) or MPTP-treated ( n = 5) mice euthanized at day 2 after the last injection. Quantification is depicted as the fold change of COX IV intensity in TH-positive neurons compared to vehicle condition. White arrows indicate representative neurons. A minimum of 36 neurons were analyzed per animal (* P < 0.05 after unpaired two-sided Student’s t -test). Error bars indicate s.e.m.

Article Snippet: Human TIM23 and TOM20 plasmids cloned into a pCMV6-XL5 vector were obtained from Origene (Origene Technologies), respectively.

Techniques: Western Blot, Injection

Journal: Cell Death & Disease

Article Title: Defective mitochondrial protein import contributes to complex I-induced mitochondrial dysfunction and neurodegeneration in Parkinson’s disease

doi: 10.1038/s41419-018-1154-0

Figure Lengend Snippet: a Representative images of TIM23 and TH immunofluorescence in ventral midbrain sections from AAV-hTIM23 mice ( n = 3). Scale bar: 150 μm. b – d Sham- and AAV-hTIM23-injected mice were treated with vehicle or MPTP. b Representative images of TH and COX IV immunofluorescence in ventral midbrain sections (Sham-Vehicle n = 5; Sham-MPTP n = 6; AAV-hTIM23-Vehicle n = 5; AAV-hTIM23-MPTP n = 6). Quantification is depicted as the fold change in COX IV intensity compared with sham-injected vehicle-treated group (* P < 0.05, *** P < 0.001 after two-way ANOVA followed by Tukey’s post hoc test). Scale bar: 50 μm. c Representative photomicrographs of TH immunohistochemistry in striatum (Sham-Vehicle n = 7; Sham-MPTP n = 5; AAV-hTIM23-Vehicle n = 8; AAV-hTIM23-MPTP n = 8). Quantification is depicted as the optical densitometry of striatal TH immunoreactivity in the different experimental groups at day 21 post MPTP (** P < 0.01 after two-way ANOVA followed by Tukey’s post hoc test). Scale bar: 500 μm. d Representative photomicrographs of TH immunohistochemistry SNpc (Sham-Vehicle n = 14; Sham-MPTP n = 10; AAV-hTIM23-Vehicle n = 7; AAV-hTIM23-MPTP n = 8). Quantification is depicted as the stereological cell counts of SNpc TH-immunoreactive neurons in the different experimental groups at day 21 post MPTP (* P < 0.05, *** P < 0.001 after two-way ANOVA followed by Tukey’s post hoc test). Scale bar: 500 μm. Error bars indicate s.e.m.

Article Snippet: Human TIM23 and TOM20 plasmids cloned into a pCMV6-XL5 vector were obtained from Origene (Origene Technologies), respectively.

Techniques: Immunofluorescence, Injection, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis

doi: 10.1038/s41419-026-08493-8

Figure Lengend Snippet: A , B Immunohistochemistry analyses of PGC-1α, Nrf1, Tim23 and Drp1 expression and Drp1 Ser616 phosphorylation in the liver sections ( A ) from human subjects and ( B ) from the HFD-Fed mice. C, E. Western blot analyses of the relative levels of PGC-1α, Nrf1, Tim23, Drp1, and Drp1 Ser616 phosphorylation to GAPDH expression in the livers of HFD-fed mice ( C , C’ ) and in the PA-treated primary hepatocytes ( E , E’ ). D , F RT-qPCR analyses of the relative levels of PGC-1α, Nrf1, Tim23 and Drp1 mRNA transcripts in the livers of HFD-fed mice ( D ) and in the PA-treated primary hepatocytes ( F ). G , H HEK293T cells were transfected with mouse Nrf1 ( G ), Tim23 ( H ) luciferase-reporter or mutant constructs, together with Ppargc1 α ( G ) or Nrf1 ( H ) or expression plasmid or the control vector, and the relative luciferase activities were measured. I , I’ , J , J’ Primary hepatocytes were transfected with negative control (NC), Nrf1 SiRNA (Si-Nrf1) or Tim23 SiRNA (Si-Tim23), leading to four groups of the Si-NC, Si-NC + PA, Si-Nrf1 and Si-Nrf1+PA or Si-NC, Si-NC + PA, Si-Tim23 and Si-Tim23+PA. Western blot analyses of the relative levels of Nrf1 and Tim23 to the control GAPDH proteins ( I , I’ ). Western blot analyses of the relative levels of Tim23 and Drp1 to the control GAPDH proteins ( J , J’ ). Data are representative images or shown as mean ± SD of each group ( n = 3–6 per group) from at least three separate experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

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).

Techniques: Immunohistochemistry, Expressing, Phospho-proteomics, Western Blot, Quantitative RT-PCR, Transfection, Luciferase, Mutagenesis, Construct, Plasmid Preparation, Control, Negative Control

Journal: Cell Death & Disease

Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis

doi: 10.1038/s41419-026-08493-8

Figure Lengend Snippet: Wild-type C57BL/6 and PGC-1α-KO mice were randomized and fed with normal chow or HFD for 16 weeks, leading to four groups of the Control, HFD, PGC-1α-KO, and PGC-1α-KO + HFD. A H&E, Oil Red O, Sirius Red, Perls’ Blue staining, and immunohistochemistry analyses of PGC-1α, Tim23, Drp1, P-Drp1 Ser616 and ACSL4 expression in liver sections. B – E The levels of serum ALT, TG, T-CHO and LDL-C. F The levels of serum irons. G The levels of hepatic GSH. H , H’ Western blot analyses of the relative levels of hepatic PGC-1α, Tim23, Drp1, P-Drp1 Ser616 , ACSL4, GPX4, TFR1, and FTH1 to the GAPDH in mice. I RT-qPCR analyses of the relative levels of Drp1, ACSL4, TFR1 and GPX4 mRNA transcripts. Data are representative images or shown as mean ± SD of each group (5–10/group) from at least three separate experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

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).

Techniques: Control, Staining, Immunohistochemistry, Expressing, Western Blot, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis

doi: 10.1038/s41419-026-08493-8

Figure Lengend Snippet: Primary hepatocytes were transfected with negative control (NC) or PGC-1α-specific siRNA (Si-PGC-1α) and treated with PA for 48 h, leading to four groups of the Si-NC, Si-NC + PA, Si-PGC-1α, and Si-PGC-1α + PA. A Oil Red O, Perls’ Blue staining and Tim23 Immunohistochemistry of hepatocytes. B , B’ Western blot analyses of the relative levels of PGC-1α, Tim23, Drp1, P-Drp1 Ser616 , ACSL4, GPX4, TFR1, and FTH1 to GAPDH expression. C RT-qPCR analyses of the relative levels of PGC-1α, Drp1, ACSL4 and TFR1 mRNA transcripts. D GSH levels. E Images of MitoTracker and MitoSOX staining. F Images of C11-BODIPY 581/591 staining. Data are representative images or expressed as the mean ± SD of each group ( n = 3) from at least three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

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).

Techniques: Transfection, Negative Control, Staining, Immunohistochemistry, Western Blot, Expressing, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis

doi: 10.1038/s41419-026-08493-8

Figure Lengend Snippet: A A diagram illustrated the study design for HFD-MASH mice treated with AAV8-PGC-1α and AAV8-NC, leading to four groups of AAV8-NC, AAV8-NC + HFD, AAV8-PGC-1α, and AAV8-PGC-1α + HFD. A’ The liver-to-body weight ratios in mice. B Macroscopic Examination, H&E, Sirius Red, Perls’ Blue, and immunohistochemistry analyses of PGC-1α, Tim23, Drp1, P-Drp1 Ser616 and ACSL4 expression in liver sections. C – F The levels of serum ALT, TG, T-CHO, and LDL-C. G , H The levels of serum and hepatic irons. I , J Hepatic GSH and MDA contents. K Transmission electron microscopy analysis of hepatocyte mitochondria. L , L’ Western blot analyses of the relative levels of hepatic PGC-1α, Tim23 Drp1, P-Drp1 Ser616 , ACSL4, GPX4, TFR1 and FTH1 to GAPDH. M RT-qPCR analyses of the relative levels of PGC-1α, Drp1, ACSL4, TFR1, GPX4, and FTH1 mRNA transcripts. Data are representative images or shown as mean ± SD of each group (5–10/group) from at least three separate experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

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).

Techniques: Immunohistochemistry, Expressing, Transmission Assay, Electron Microscopy, Western Blot, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis

doi: 10.1038/s41419-026-08493-8

Figure Lengend Snippet: Primary hepatocytes were transfected with negative control (NC) or PGC-1α CRISPR activation plasmid (PGC-1α CRISPR ACT) and treated with PA, leading to four groups of the Act-NC, Act-NC + PA, Act-PGC-1α and Act-PGC-1α + PA. A Oil Red O, Perls’ Blue staining and Tim23 Immunohistochemistry. B , B ’ Western blot analyses of the relative levels of hepatic PGC-1α, Tim23, Drp1, P-Drp1 Ser616 , ACSL4, GPX4 and FTH1 to GAPDH. C RT-qPCR analyses of the relative levels of PGC-1α, Drp1, ACSL4, GPX4, TFR1 and FTH1 mRNA transcripts. D The GSH levels. E Images of MitoTracker and MitoSOX staining. F Images of C11-BODIPY 581/591 staining. Data are representative images or expressed as the mean ± SD of each group ( n = 3) from at least three independent treatments. * P < 0.05, ** P < 0.01, *** P < 0.001.

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).

Techniques: Transfection, Negative Control, CRISPR, Activation Assay, Plasmid Preparation, Staining, Immunohistochemistry, Western Blot, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: PGC-1α protects against MASH via Tim23-dependent inhibition of DRP1-mediated ferroptosis

doi: 10.1038/s41419-026-08493-8

Figure Lengend Snippet: Primary hepatocytes were transfected with negative control (NC) or Tim23 SiRNA (Si-Tim23), leading to four groups of the Si-NC, Si-NC + PA, Si-Tim23 and Si-Tim23+PA. A Oil Red O and Perls’ Blue staining of hepatocytes. B , B ’ Western blot analyses of the relative levels of Tim23, Drp1, ACSL4, GPX4 and FTH1 to the GAPDH expression in primary hepatocytes. C Immunofluorescence analyses of Drp1 and COXIV co-localization. D Mitochondrial membrane potential (MMP) was determined by JC-1 staining. E Images of C11-BODIPY 581/591 staining in primary hepatocytes. Data are representative images or expressed as the mean ± SD of each group ( n = 3) from at least three independent treatments. * P < 0.05, ** P < 0.01, *** P < 0.001.

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).

Techniques: Transfection, Negative Control, Staining, Western Blot, Expressing, Immunofluorescence, Membrane

Journal: Nucleic acids research

Article Title: DNA damage response regulator ATR licenses PINK1-mediated mitophagy.

doi: 10.1093/nar/gkaf178

Figure Lengend Snippet: Figure 3. ATR docks at mitochondria by direct interaction with PINK1 and TOM / TIM. ( A ) PLAs of ATR interaction with PINK1, TIM23, and TOM40 in U2OS cells without (shNC) or with ATR-KD (shATR). Left panel: Representative images of PLA-positive signals (red foci) after co-staining with mitochondrial marker MitoTracker. Yellow arrows indicate PLA foci in mitochondria. Right panel: Quantification of PLA-positive foci from > 200 cells scored. Statistics were performed using a two-way ANO V A with a Tukey’s multiple comparisons test. n = 3. ( B ) Co-IPs by the FLAG antibody in FLAG-ATR and HA-TOM40 transfected HEK239T cells f ollo w ed b y w estern blot analy sis of the indicated proteins. β-actin w as used to control protein loading. n = 3. ( C ) Co-IPs by the FLAG antibody in FLAG-ATR and HA-TIM23 transfected HCT116 cells f ollo w ed b y w estern blot analy sis of the indicated proteins. β-actin was used to control protein loading. n = 3. ( D ) HCT116 cells with targeted siRNA against ATR (siATR), PINK1 (siPINK1), HSP90 α (siHSP90), TIM23 (siTIM23), and TOM7 (siTOM7) are analyzed by Western blotting for the indicated proteins in mitochondrial fractionations. ATP5A was used to control protein loading. n = 3. ( E ) 1D BN–PAGE analysis of ATR, TOM22, and PINK1 from the purified mitochondrial fraction of N2A cells. Under native conditions, ATR, TOM22, and PINK1 migrate at the same speed in native PAGE gel. After heat treatment, denatured samples were pulled down by an anti-ATR ab. The native and denatured samples were blotted with the indicated antibodies, which detect the respective proteins in denatured gels (lane “heat denaturing +”). n = 3.

Article Snippet: Scrambled siRNAs (siCtl: equal mixture of sc-37007, c-44230, and sc-44231) and targeted siRNAs against ATR siATR#1: sc-29763), PINK1 (siPINK1#1: sc-44598), TIM23 sc-44155), TOM7 (sc-89537), and HSP90 (sc-35608) were urchased from Santa Cruz Biotechnologies, and (siATR#2: HU040341-50UG) (siATR mix: equal mixture of siATR#1 nd #2) and (siPINK1#2: EHU057101-5OUG) (siPINK1 mix: qual mixture of siPINK1#1 and #2) from Sigma–Aldrich.

Techniques: Staining, Marker, Transfection, Control, Western Blot, Purification, Clear Native PAGE

Journal: EMBO Molecular Medicine

Article Title: Parkin is a disease modifier in the mutant SOD 1 mouse model of ALS

doi: 10.15252/emmm.201808888

Figure Lengend Snippet: A, B Representative Western blot (A) and quantification (B) of p62 associated with mitochondria from spinal cords. Protein levels are normalized by subunit ATPase β of mitochondria (Complex V). Mitochondrial p62 levels are increased at 120 days (symptomatic stage) in SOD1‐G93A spinal cord relative to Non Tg and wild‐type SOD1 (wtSOD1). Results are expressed as mean ± SEM relative to Non Tg at 30 days; n = 8 mice (four males and four females). At 120 days, ** P = 0.0018 (Non Tg vs. SOD1‐G93A) and ** P = 0.0011 (SOD1‐G93A vs. wtSOD1) by paired one‐way ANOVA with Tukey's correction. No other statistically significant differences were found (paired Friedman's test with Dunn's correction at 30 days and paired one‐way ANOVA with Tukey's correction at 60 and 90 days). C, D Representative Western blot (C) and quantification (D) of Tim23 in homogenates from spinal cords using β‐actin as normalizer. Results are expressed as mean ± SEM and relative to Non Tg at 30 days; n = 6 mice (three males and three females). *** P = 0.0002 (Non Tg vs. SOD1‐G93A at 120 days) by paired one‐way ANOVA with Tukey's correction. No other statistically significant differences were found (paired Friedman's test with Dunn's correction at 30 days and paired one‐way ANOVA with Tukey's correction at 60 and 90 days). Tim23 is decreased in SOD1‐G93A spinal cord relative to Non Tg at 120 days. E, F Western blot (E) and quantification (F) of COXI in homogenates from spinal cords. Protein levels are normalized by β‐actin. Results are expressed as mean ± SEM and relative to Non Tg at 30 days; n = 6 mice (three males and three females). * P = 0.017 (Non Tg vs. SOD1‐G93A at 120 days) by paired Friedman's test with Dunn's correction. No other statistically significant differences were found (paired Friedman's test with Dunn's correction at 30 and 60 days and paired one‐way ANOVA with Tukey's correction at 90 days). COX1 is decreased in SOD1‐G93A spinal cord relative to Non Tg at 120 days. G qPCR (fold change) of PGC1α mRNA normalized by β‐actin mRNA. Results are expressed as mean ± SEM and fold change of Non Tg at 30 days; n = 6 (three males and three females) for 30 and 120 days, n = 4 (two males and two females) for 60 days and n = 5 (three males and two females) for 90 days. * P = 0.035 at 90 days and *** P = 0.0007 at 120 days. Paired Student's t ‐test (for 60, 90, and 120 days) and paired Wilcoxon's test (for 30 days) were used for comparisons. Source data are available online for this figure.

Article Snippet: The following antibodies were used in Western blot (additional information can be found in ): Parkin (clone PRK8, 1:1,000, cat. # sc‐32282, Santa Cruz Biotechnology; Dallas, TX), β‐actin (clone AC‐74, 1:1,000–1:5,000, cat. # A5316, Sigma), p62 (clone 2C11, 1:1,000, cat. # H00008878‐M01, Novus Biologicals; Littleton, CO), optineurin (OPTN, 1:1,000, cat. # 10837‐1‐AP, Proteintech; Rosemont, IL), Complex V (ATP synthase beta, clone 3D5AB1, 1:1,000–1:5,000, cat. # A21351, Thermo Fisher Scientific; Waltham, MA), Tim23 (clone 32/Tim23, 1:1,000, cat. # 611222, BD Transduction Labs; San Jose, CA), COX1 (Complex IV, subunit 1, clone 1D6E1A8, 1:2,000, cat. #459600, Thermo Fisher Scientific), SOD1 (1:1,000, cat. # 574597, EMD Millipore; Billerica, MA), VCP (valosin‐containing protein, clone 5, 1:1,000, cat. # MA3‐004, Thermo Scientific; Rockford, IL), LC3 (microtubule‐associated protein 1A/1B‐light chain 3B, LC3B, 1:1,000, cat. # L7543, Sigma), March5 (membrane‐associated ring‐CH‐type finger 5, 1:1,000, cat. # PA5‐25584, Thermo Fisher Scientific), Mul1 (mitochondrial E3‐ubiquitin protein ligase 1 1:1,000, cat. # HPA017681, Sigma), Miro1 (mitochondrial Rho GTPase 1, clone 4H4, 1:1,000, cat. # H00055288‐M01, Novus Biologicals), mitofusin 2 (Mfn2, 1:1,000, cat. # M6319, Sigma), PARIS (parkin‐interacting substrate (ZNF746), clone N196/16, 1:1,000, cat. # 75‐195, NeuroMabs and Antibodies Inc.; Davis, CA), ubiquitin Lys48‐specific (clone Apu2, 1:1,000, cat. # 05‐1307, EMD Millipore), ubiquitin Lys63‐specific (clone Apu3, 1:1,000, cat. # 05‐1308, EMD Millipore), and horseradish‐peroxidase (HRP) goat anti‐rabbit, goat anti‐mouse, and donkey anti‐sheep secondary antibodies (1:10,000, Jackson ImmunoResearch; West Groove, PA).

Techniques: Western Blot

Journal: EMBO Molecular Medicine

Article Title: Parkin is a disease modifier in the mutant SOD 1 mouse model of ALS

doi: 10.15252/emmm.201808888

Figure Lengend Snippet: A, B Western blots of COXI (A) and Tim23 (B) in spinal cord homogenates at 130 days. C Quantification of COXI at 130 days with β‐actin as a normalizer. Results are expressed as mean ± SEM and as percent of Non Tg; n = 8 (four males and four females) mice per group. No statistically significant differences were found between G93A and PKO/G93A ( P = 0.493 by paired Student's t ‐test). D Quantification of Tim23 at 130 days, using β‐actin as loading control, showed decreased levels of Tim23 in G93A mice. Results are expressed as mean ± SEM and as a percent of Non Tg; n = 8 (four males and four females) mice per group. No statistically significant differences were found between G93A and PKO/G93A ( P = 0.921 by paired Student's t ‐test). * P = 0.035 (Non Tg Vs. G93A) by paired Friedman's test with Dunn's correction. No other statistically significant differences were found. E, F Representative Western blots of COXI (E) and Tim23 (F) in disease end‐stage homogenates. G Quantification of COXI at disease end stage using β‐actin as loading control. Results are expressed as mean ± SEM and as percent of Non Tg; n = 5 (three males and two females) mice per group. * P = 0.047 for (G93A and PKO/G93A) by paired Student's t ‐test; ** P = 0.0018 by paired Friedman's test with Dunn's correction (Non Tg vs. G93A). H Quantification of Tim23 with β‐actin as normalizer at end stage showed that Parkin knockout can alleviate the turnover of mitochondrial proteins in G93A spinal cords. Results are expressed as mean ± SEM and as percent of Non Tg; n = 8 (four males and four females) mice per group. No statistically significant differences were found between G93A and PKO/G93A ( P = 0.190 by paired Student's t ‐test). * P = 0.037 (for PKO vs. PKO/G93A) by paired Friedman's test with Dunn's correction. No other statistically significant differences were found. Source data are available online for this figure.

Article Snippet: The following antibodies were used in Western blot (additional information can be found in ): Parkin (clone PRK8, 1:1,000, cat. # sc‐32282, Santa Cruz Biotechnology; Dallas, TX), β‐actin (clone AC‐74, 1:1,000–1:5,000, cat. # A5316, Sigma), p62 (clone 2C11, 1:1,000, cat. # H00008878‐M01, Novus Biologicals; Littleton, CO), optineurin (OPTN, 1:1,000, cat. # 10837‐1‐AP, Proteintech; Rosemont, IL), Complex V (ATP synthase beta, clone 3D5AB1, 1:1,000–1:5,000, cat. # A21351, Thermo Fisher Scientific; Waltham, MA), Tim23 (clone 32/Tim23, 1:1,000, cat. # 611222, BD Transduction Labs; San Jose, CA), COX1 (Complex IV, subunit 1, clone 1D6E1A8, 1:2,000, cat. #459600, Thermo Fisher Scientific), SOD1 (1:1,000, cat. # 574597, EMD Millipore; Billerica, MA), VCP (valosin‐containing protein, clone 5, 1:1,000, cat. # MA3‐004, Thermo Scientific; Rockford, IL), LC3 (microtubule‐associated protein 1A/1B‐light chain 3B, LC3B, 1:1,000, cat. # L7543, Sigma), March5 (membrane‐associated ring‐CH‐type finger 5, 1:1,000, cat. # PA5‐25584, Thermo Fisher Scientific), Mul1 (mitochondrial E3‐ubiquitin protein ligase 1 1:1,000, cat. # HPA017681, Sigma), Miro1 (mitochondrial Rho GTPase 1, clone 4H4, 1:1,000, cat. # H00055288‐M01, Novus Biologicals), mitofusin 2 (Mfn2, 1:1,000, cat. # M6319, Sigma), PARIS (parkin‐interacting substrate (ZNF746), clone N196/16, 1:1,000, cat. # 75‐195, NeuroMabs and Antibodies Inc.; Davis, CA), ubiquitin Lys48‐specific (clone Apu2, 1:1,000, cat. # 05‐1307, EMD Millipore), ubiquitin Lys63‐specific (clone Apu3, 1:1,000, cat. # 05‐1308, EMD Millipore), and horseradish‐peroxidase (HRP) goat anti‐rabbit, goat anti‐mouse, and donkey anti‐sheep secondary antibodies (1:10,000, Jackson ImmunoResearch; West Groove, PA).

Techniques: Western Blot, Knock-Out

Journal: Antioxidants & Redox Signaling

Article Title: Cysteine Deprivation Targets Ovarian Clear Cell Carcinoma Via Oxidative Stress and Iron−Sulfur Cluster Biogenesis Deficit

doi: 10.1089/ars.2019.7850

Figure Lengend Snippet: Cysteine deprivation damages mitochondria of respiring OCCC via Fe–S cluster biogenesis deficit. (A) Mitochondria membrane potential in KOC-7C 3 days post-treatment was visualized using TMRM fluorescent dye. FCCP-treated cells (50 μ M for an hour) served as a negative control. Scale bar, 50 μm. (B) Representative Western blot of cyt c upon mitochondria isolation in KOC-7C 3 days post-treatment. TIM23 was used as a mitochondria-specific marker. GAPDH was used as a cytosolic marker. ( C ) OCR of nontreated (NT) versus SAS + PAG-treated KOC-7C cells (SP). Electron transport chain inhibitors oligomycin (1 μ M ), FCCP (0.5 μ M ), and Rot/AA (1 μ M ) were added at the indicated time points (—). (D) ECAR of nontreated (NT) versus SAS + PAG-treated KOC-7C cells (SP). Glucose (10 m M ), oligomycin (2 μ M ), and 2-DG (50 m M ) were added at the indicated time points (—). (E) ATP production in KOC-7C with and without 1-h pretreatment with NAC (5 m M ). Data represent the means ± SD ( n = 3); ***p < 0.001. (F) Volcano plot showing differentially expressed proteins between control and cysteine-deprived KOC-7C. Succinate dehydrogenase B (SDHB), ATP-binding cassette subfamily E member 1 (ABCE1), molybdenum cofactor sulfurase (MOCOS), CDGSH iron−sulfur domain-containing protein 1 (C10ORF70), ferredoxin reductase (FDXR), iron-responsive element-binding protein (IREB2), cytochrome c 1 (CYC1), mitochondrial fission regulator 1 like (MTFR1L), ATPase H+ transporting accessory protein 1 (ATP6AP1), mannose-6-phosphate receptor (M6PR), frataxin (FXN), nuclear receptor coactivator 4 (NCOA4), glutaredoxin 2 (GLRX2). ( n = 3). (G) Relative expressions of Fe−S cluster proteins, as shown in volcano plot , in KOC-7C 3 days post-treatment. (H) ACO1 enzymatic activity in KOC-7C 3 days post-treatment with and without 1-h pretreatment with 5 m M NAC. DFO-treated cells (5 μ M for 3 days) served as a negative control. Data represent the means ± SD ( n = 3); **p < 0.01, ***p < 0.001. ACO1, aconitase 1; Fe−S, iron−sulfur; TMRM, tetramethyl rhodamine. Color images are available online.

Article Snippet: The primary antibodies used in this study were as follows: Mouse α-CTH (Novus; Cat. #H00001491-M01), Rabbit α-xCT/SLC7A11 (Cell Signaling Technology [CST]; Cat. #12691), Rabbit α-cleaved caspase 3 (CST; Cat. #9661), Rabbit α-caspase 3 (CST; Cat. #9662), Rabbit α-cleaved PARP (CST; Cat. #9541), Rabbit cytochrome c (CST; Cat. #4280), Mouse α-TIM23 (Biomed Diagnostics; Cat. #611222), Rabbit α-ACO1 (CST; Cat. #20272), Rabbit α-IREB2 (D6E6W, Cat. #37135; CST), Rabbit α-ferritin (Abcam; Cat. #ab75973), Rabbit α-TFRC (clone D7S5Z, Cat. #13208; CST), Rabbit α-phosphorylated S6 (CST; Cat. #4858), Rabbit α-phosphorylated p70S6 kinase (CST; Cat. #9234), Rabbit α-p70S6 kinase (CST; Cat. #2708), Rabbit α-LC3B (CST; Cat. #4108), and Rabbit α-GAPDH (Santa Cruz Biotechnology; Cat. #sc-25778).

Techniques: Negative Control, Western Blot, Isolation, Marker, Binding Assay, Activity Assay

Journal: Antioxidants & Redox Signaling

Article Title: Cysteine Deprivation Targets Ovarian Clear Cell Carcinoma Via Oxidative Stress and Iron−Sulfur Cluster Biogenesis Deficit

doi: 10.1089/ars.2019.7850

Figure Lengend Snippet: Cysteine deprivation promotes iron buildup in mitochondria. (A) Daily LIP in KOC-7C. 100 μ M DFO was used to dequench 100 n M calcein-AM fluorescence overnight. Data represent the means ± SD ( n = 3); ***p < 0.001. (B) Daily mitoferrin 2 ( MFRN2 also known as SLC25A28 ) gene expression in KOC-7C as measured by quantitative real-time polymerase chain reaction. Data represent the means ± SD ( n = 3); ***p < 0.001. (C) Mitochondria iron (mito. iron), detected by SQX.RPA1 in KOC-7C 3 days post-treatment. One hundred micromolar DFO or BIP were used to dequench 1 μ M RPA fluorescence overnight. Data represent the means ± SD ( n = 3); ***p < 0.001. (D) Representative Western blot of ferritin upon mitochondria isolation in KOC-7C 3 days post-treatment with combination of SAS and PAG. TIM23 was used as a mitochondria-specific marker. GAPDH was used as a cytosolic marker. (E) Mitochondria superoxide in KOC-7C 3 days post-treatment was visualized by MitoSOX fluorescent dye. AA-treated cells (50 μ M for an hour) served as a positive control. Scale bar, 50 μm. BIP, 2,2′-bipyridyl; LIP, labile iron pool; RPA, rhodamine B-[1,10-phenanthrolin-5-yl)-aminocarbonyl] benzylester. Color images are available online.

Article Snippet: The primary antibodies used in this study were as follows: Mouse α-CTH (Novus; Cat. #H00001491-M01), Rabbit α-xCT/SLC7A11 (Cell Signaling Technology [CST]; Cat. #12691), Rabbit α-cleaved caspase 3 (CST; Cat. #9661), Rabbit α-caspase 3 (CST; Cat. #9662), Rabbit α-cleaved PARP (CST; Cat. #9541), Rabbit cytochrome c (CST; Cat. #4280), Mouse α-TIM23 (Biomed Diagnostics; Cat. #611222), Rabbit α-ACO1 (CST; Cat. #20272), Rabbit α-IREB2 (D6E6W, Cat. #37135; CST), Rabbit α-ferritin (Abcam; Cat. #ab75973), Rabbit α-TFRC (clone D7S5Z, Cat. #13208; CST), Rabbit α-phosphorylated S6 (CST; Cat. #4858), Rabbit α-phosphorylated p70S6 kinase (CST; Cat. #9234), Rabbit α-p70S6 kinase (CST; Cat. #2708), Rabbit α-LC3B (CST; Cat. #4108), and Rabbit α-GAPDH (Santa Cruz Biotechnology; Cat. #sc-25778).

Techniques: Fluorescence, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Isolation, Marker, Positive Control