Journal: Oxidative medicine and cellular longevity

Article Title: Mitochondrial-Targeting Antioxidant SS-31 Suppresses Airway Inflammation and Oxidative Stress Induced by Cigarette Smoke.

doi: 10.1155/2021/6644238

Figure Lengend Snippet: Figure 6: SS-31 prevents mitochondrial dysfunction in CS-induced mouse lung. (a–d) Protein levels of OPA1, MFF (a, b), and cytochrome c (c, d) in lungs were evaluated by western blot following exposure to CS with or without SS-31 pretreatment. GAPDH or COX IV was used as a loading control (n = 3 per group). ∗∗P < 0:01 and ∗∗∗P < 0:001 vs. control; #P < 0:05 vs CS. Abbreviations: COX IV: cytochrome c oxidase IV; CS: cigarette smoke; Cyt c: cytochrome c; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; MFF: mitochondrial fission factor; mit: mitochondrion; OPA1: optic atrophy 1; SS-31 (H): high-dose SS-31 (5 mg/kg); SS-31 (L): low-dose SS-31 (2.5 mg/kg).

Article Snippet: After blocking with 5% bovine serum albumin in Trisbuffered saline at room temperature for 1 h, the membrane was incubated overnight at 4°C with antibodies against optic atrophy (OPA) 1 (mouse antibody from Cell Signaling Technology; human antibody from Proteintech, Rosemont, IL, USA); mitochondrial fission factor (MFF) (mouse antibody from Cell Signaling Technology; human antibody from Proteintech); and extracellular signal-regulated kinase (ERK), phosphorylated- (P-) ERK, P38, P-P38, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (all from Cell Signaling Technology).

Techniques: Western Blot, Control

Journal: Oxidative medicine and cellular longevity

Article Title: Mitochondrial-Targeting Antioxidant SS-31 Suppresses Airway Inflammation and Oxidative Stress Induced by Cigarette Smoke.

doi: 10.1155/2021/6644238

Figure Lengend Snippet: Figure 11: SS-31 protects against CSE-induced mitochondrial dysfunction via suppression of MAPK signaling. (a, b) Protein levels of OPA1 and MFF detected by western blot in BEAS-2B cells (n = 3 per group). ∗∗P < 0:01, ∗∗∗P < 0:001, and ∗∗∗∗P < 0:0001 vs. control; #P < 0:05 and ##P < 0:01 vs. ani+CSE; &P < 0:05 vs. CSE. Abbreviations: ani: anisomycin; CSE: cigarette smoke extract; MFF: mitochondrial fission factor; OPA1: optic atrophy 1.

Article Snippet: After blocking with 5% bovine serum albumin in Trisbuffered saline at room temperature for 1 h, the membrane was incubated overnight at 4°C with antibodies against optic atrophy (OPA) 1 (mouse antibody from Cell Signaling Technology; human antibody from Proteintech, Rosemont, IL, USA); mitochondrial fission factor (MFF) (mouse antibody from Cell Signaling Technology; human antibody from Proteintech); and extracellular signal-regulated kinase (ERK), phosphorylated- (P-) ERK, P38, P-P38, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (all from Cell Signaling Technology).

Techniques: Western Blot, Control

Journal: American journal of physiology. Heart and circulatory physiology

Article Title: High-fat, low-carbohydrate diet promotes arrhythmic death and increases myocardial ischemia-reperfusion injury in rats.

doi: 10.1152/ajpheart.00058.2014

Figure Lengend Snippet: Fig. 6. Effect of HFLCD on key mitochondrial fusion and fission regulatory proteins. A: real-time PCR measurement of mitofusin (Mfn)1, Mfn2, optic atrophy 1 (Opa1), fission1 (Fis1), and dynamin-related protein 1 (Drp1) transcript in hearts following I/R. B: Western blot analyses of Mfn1, Mfn2, Opa1, Fis1 and Drp1 from hearts following I/R. Values are means SE, n 4 to 5 each group, *P 0.05 vs. CONT.

Article Snippet: Western blots were conducted using commercial antibodies: carnitine palmitoyltransferase (CPT1B, CPT2), uncoupling proteins (UCP2, UCP3) (Alpha Diagnostic, San Antonio, TX); ATP synthase (MitoSciences, Eugene, OR); Mfn1, Opa1, GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA); Fis1, Drp1 (Thermo Fisher Scientific, Rockford, IL); Mfn2 (Abcam, Cambridge, MA); STIM1 (Cell Signaling Technology, Danvers, MA); and Orai1, Orai3 (ProSci, Poway, CA).

Techniques: Real-time Polymerase Chain Reaction, Western Blot

Journal: Frontiers in pharmacology

Article Title: Buyang Huanwu Decoction alleviates cerebral ischemic injury through modulating caveolin-1-mediated mitochondrial quality control.

doi: 10.3389/fphar.2023.1137609

Figure Lengend Snippet: FIGURE 5 BHD maintains mitochondrial dynamic balance in MCAO mice through Cav-1. (A–E) Western blot and quantitative analysis of DRP1, FIS1, MFN2 and OPA1 (n = 3). (F–I) RT‒qPCR and quantitative analysis of DRP1, FIS1, MFN2 and OPA1 (n = 6). All data are presented as the mean ± SD. **p < 0.01 vs. syngeneic sham group, #p < 0.05 and ##p < 0.01 vs. syngeneic model group, ▲p < 0.05 and ▲▲p < 0.01 vs. WT model group, ◇p < 0.05 and ◇◇p < 0.01 vs. WT BHD group.

Article Snippet: NeuN antibody (Proteintech, 26975-1-AP,Wuhan, China), dynamin 1-like (DRP1) antibody (Boster, A00556-2,Wuhan, Frontiers in Pharmacology frontiersin.org02 China), fission 1 (FIS1) (Boster, A01932-2,Wuhan, China), mitofusin 2 (MFN2) (Boster, BM4906,Wuhan, China), optic atrophy 1 (OPA1) (Boster, PB0773,Wuhan, China), PTEN induced putative kinase 1 (PINK1) (Proteintech, 23274-1- AP,Wuhan, China), PARKIN (Boster, PB9307,Wuhan, China), Beclin 1 (Boster, PB9076,Wuhan, China), microtubule-associated protein 1 light chain 3 beta (LC3) antibody (Proteintech, 14600-1- AP,Wuhan, China), sirtuin 1 (SIRT1) antibody (Boster, A000181,Wuhan, China), peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC-1α)Antibody (Proteintech, 66369-1-Ig,Wuhan, China), adenosine triphosphate (ATP) assay kit (Jiancheng, A095-1, Nanjing, China), ATPase assay kit (Jiancheng, A016-1, Nanjing, China), RNA Extraction Kit (Beyotime, R0026, Beijing, China), First Strand cDNA Synthesis Kit (Beijing Beyotime, D7178), Universal Genomic DNA Purification Mini Spin Kit (Beyotime, D0063, Beijing, China), SYBR Green qPCR Mix (Beyotime, product number D7265, Beijing, China), the PCR primers were synthesized by Sangon (Shanghai, China), citrate synthase (CS) assay kit (Solarbio, BC1060, Beijing, China), Nicotinamide adenine dinucleotide (NADH)- coenzyme Q reductase assay kit (Solarbio, BC0510, Beijing, China), succinate-coenzyme Q assay kit (Solarbio, BC3230, Beijing, China), coenzyme Q-cytochrome C assay kit (Solarbio, BC3240, Beijing, China), cytochrome C oxidase assay kit (Solarbio, BC0940, Beijing, China).

Techniques: Western Blot

Journal: Cells

Article Title: A Role for PGC-1a in the Control of Abnormal Mitochondrial Dynamics in Alzheimer’s Disease

doi: 10.3390/cells11182849

Figure Lengend Snippet: MutAPP triggers dynamic imbalance of mitochondrial fission and fusion. N 2 A cells were transfected with pCDNA or APPswe plasmid for 48 h. ( A ) EGFP-labeled APPswe was successfully overexpressed in N 2 A cells. Expression patterns and qualification of the fusion proteins, including ( B , D ) OPA1, ( F , I ) MFN1 and ( G , J ) MFN2, as well as the fission proteins, including ( L , N ) DRP1 and ( P , R ) FIS1 were studied with Western blot. Expression patterns and qualification of ( C , E ) OPA1, ( H , K ) MFN2, ( M , O ) DRP1 and ( Q , S ) FIS1 from the parietal cortex samples of 2×Tg-AD mice were also examined with immunohischemistry. Scale bars = 100 μm. ( T ) Representative electron microscope images showing stages of mitochondria in cortical neurons of WT and APP/PS1 animals. Scale bar = 0.5 μm. Yellow stars: the fissive stage of mitochondria. Significance levels were set at ** p < 0.01, *** p < 0.001 for noted differences between pCDNA and APPswe groups and WT and APP/PS1 groups. Tubulin was used as the loading control.

Article Snippet: Primary antibodies against EGFP (1:1000, Beyotime, cat # AG281, Shanghai, China), Flag (1:1000, abm, cat # G188, Zhenjiang, China), OPA1 (1:1000, Boster, cat # PB0773, Wuhan, China), MFN1 (1:1000, Boster, cat # PB0263, Wuhan, China), MFN2 (1:800, Bioss, cat # bs-23685R, Beijing, China), DRP1 (1:1000, Wanleibio, cat # WL03028, Shenyang, China), FIS1 (1:1000, Boster, cat # A01932-2, Wuhan, China), BAX (1:5000, Abcam, cat # ab32503, Cambridge, MA, USA), Bcl-2 (1:2000, Abcam, cat # ab182858, Cambridge, MA, USA), PGC-1a (1:1000, Bioss, cat # bs-1832R, Beijing, China), KIF5A Flag (diluted 1:1000, Bioword, cat # BS71526, Nanjing, China), KIF5B (diluted 1:1000, Wanleibio, cat # WL04906, Shenyang, China), and the horseradish peroxidase-linked antibodies (1:5000, Beyotime, goat anti mouse IgG, cat # A0216 and goat anti rabbit IgG, cat # A0208, Shanghai, China) were used to probe these blots.

Techniques: Transfection, Plasmid Preparation, Labeling, Expressing, Western Blot, Microscopy

Journal: Cells

Article Title: A Role for PGC-1a in the Control of Abnormal Mitochondrial Dynamics in Alzheimer’s Disease

doi: 10.3390/cells11182849

Figure Lengend Snippet: PGC-1α rescues mutAPP-triggered dynamic imbalance of mitochondrial fission and fusion. N 2 A cells were transfected with pEnCMV / Pgc-1alpha plasmid and plasmid-encoding APPswe for 48 h. ( A ) Flag-labeled PGC-1α and EGFP-labeled APPswe were successfully overexpressed in N 2 A cells. Expression patterns and qualification of the fusion proteins, including ( B , D ) OPA1, ( F , I ) MFN1 and ( G , J ) MFN2, as well as the fission proteins, including ( L , N ) DRP1 and ( P , R ) FIS1, were studied with Western blot. Expression patterns and qualification of ( C , E ) OPA1, ( H , K ) MFN2, ( M , O ) DRP1 and ( Q , S ) FIS1 from the parietal cortex samples of 2×Tg-AD mice treated with Vector/ Pgc-1alpha were also examined with immunohistochemistry. Scale bars = 100 μm. ( T ) Representative electron microscopes showing stage of mitochondria in cortical neurons of AAV-Vector- and AAV- Pgc-1alpha treated APP/PS1 mice. Scale bar = 0.5 μm. Yellow stars: the fissive stage of mitochondria. Significance levels were set at ** p < 0.01, *** p < 0.001 for noted differences between APPswe + pEnCMV and APPswe + Pgc-1alpha groups, or APP/PS1 + Vector and APP/PS1 + Pgc-1alpha groups. Tubulin was used as the loading control.

Article Snippet: Primary antibodies against EGFP (1:1000, Beyotime, cat # AG281, Shanghai, China), Flag (1:1000, abm, cat # G188, Zhenjiang, China), OPA1 (1:1000, Boster, cat # PB0773, Wuhan, China), MFN1 (1:1000, Boster, cat # PB0263, Wuhan, China), MFN2 (1:800, Bioss, cat # bs-23685R, Beijing, China), DRP1 (1:1000, Wanleibio, cat # WL03028, Shenyang, China), FIS1 (1:1000, Boster, cat # A01932-2, Wuhan, China), BAX (1:5000, Abcam, cat # ab32503, Cambridge, MA, USA), Bcl-2 (1:2000, Abcam, cat # ab182858, Cambridge, MA, USA), PGC-1a (1:1000, Bioss, cat # bs-1832R, Beijing, China), KIF5A Flag (diluted 1:1000, Bioword, cat # BS71526, Nanjing, China), KIF5B (diluted 1:1000, Wanleibio, cat # WL04906, Shenyang, China), and the horseradish peroxidase-linked antibodies (1:5000, Beyotime, goat anti mouse IgG, cat # A0216 and goat anti rabbit IgG, cat # A0208, Shanghai, China) were used to probe these blots.

Techniques: Transfection, Plasmid Preparation, Labeling, Expressing, Western Blot, Immunohistochemistry

Journal: Scientific Reports

Article Title: 7,8-Dihydroxyflavone improves neuropathological changes in the brain of Tg26 mice, a model for HIV-associated neurocognitive disorder

doi: 10.1038/s41598-021-97220-8

Figure Lengend Snippet: List of primary antibodies.

Article Snippet: Anti FIS-1 , Mitochondrial fission 1 protein , Genetex, Irvine, CA, USA , 1:200 (IHC).

Techniques: Concentration Assay, Marker

Journal: Cardiovascular Research

Article Title: Mitochondrial targeted peptides preserve mitochondrial organization and decrease reversible myocardial changes in early swine metabolic syndrome

doi: 10.1093/cvr/cvx245

Figure Lengend Snippet: Mitoprotection restored subsarcolemal mitochondrial density. (A) Representative transmission electron microscopy images of cross-sectioned cardiomyocytes showing distribution of mitochondria. (B) The number of subsarcolemal mitochondria (arrows) was lower in MetS compared to Lean, but increased in MetS + ELAM (n = 6/group). (C) Mitochondrial area and matrix density did not differ among the groups (n = 6/group). *P < 0.05 vs. lean, †P < 0.05 vs. MetS + ELAM.

Article Snippet: Mitochondrial dynamics was assessed by the expression of the fusion markers mitofusin (MFN)-1 (Abnova; Cat# H00055669-P01) and optic atrophy protein (OPA)-1 (Cat# 30573; Santa Cruz Biotechnology Inc., Dallas, TX, USA), and the fission markers mitochondrial fission factor (MFF; Abnova; Cat# {"type":"entrez-protein","attrs":{"text":"PAB20518","term_id":"1236633578","term_text":"PAB20518"}} PAB20518 ) and dynamin-related protein (DRP)-1 (Cell Signaing; Cat# 8570).

Techniques: Transmission Assay, Electron Microscopy

Journal: Cardiovascular Research

Article Title: Mitochondrial targeted peptides preserve mitochondrial organization and decrease reversible myocardial changes in early swine metabolic syndrome

doi: 10.1093/cvr/cvx245

Figure Lengend Snippet: ELAM improved mitochondrial function Dand redox balance. (A) In isolated myocardial mitochondria, ATP levels were comparable in all groups, yet ADP levels were higher in MetS and normalized in MetS + ELAM, as was ATP/ADP ratio (n = 6/group). (B) COX-IV activity decreased in MetS compared to lean, but normalized in ELAM-treated pigs. (C) H2O2 production increased in MetS but normalized in MetS + ELAM. (D) GSH/GSSG and NADPH/NADP ratio decreased in MetS, but normalized in ELAM-treated pigs (n = 6/group). *P < 0.05 vs. lean, †P < 0.05 vs. MetS + ELAM .

Article Snippet: Mitochondrial dynamics was assessed by the expression of the fusion markers mitofusin (MFN)-1 (Abnova; Cat# H00055669-P01) and optic atrophy protein (OPA)-1 (Cat# 30573; Santa Cruz Biotechnology Inc., Dallas, TX, USA), and the fission markers mitochondrial fission factor (MFF; Abnova; Cat# {"type":"entrez-protein","attrs":{"text":"PAB20518","term_id":"1236633578","term_text":"PAB20518"}} PAB20518 ) and dynamin-related protein (DRP)-1 (Cell Signaing; Cat# 8570).

Techniques: Isolation, Activity Assay

Journal: Cardiovascular Research

Article Title: Mitochondrial targeted peptides preserve mitochondrial organization and decrease reversible myocardial changes in early swine metabolic syndrome

doi: 10.1093/cvr/cvx245

Figure Lengend Snippet: MetS altered mitochondria-sarcoplasmic reticulum interaction. (A) Representative myocardial double immunofluorescence staining of the mitochondrial voltage-dependent anion channel (VDAC) and the sarcoplasmic-reticulum marker ryanodine receptor (RYR). Although total VDAC and RYR fluorescence did not differ among the groups, their overlap coefficient (M1) decreased in MetS, but improved in MetS + ELAM (n = 6/group). (B) Myocardial expression of MCU decreased in MetS, but was restored in MetS + ELAM (n = 6/group). *P < 0.05 vs. lean, †P < 0.05 vs. MetS + ELAM.

Article Snippet: Mitochondrial dynamics was assessed by the expression of the fusion markers mitofusin (MFN)-1 (Abnova; Cat# H00055669-P01) and optic atrophy protein (OPA)-1 (Cat# 30573; Santa Cruz Biotechnology Inc., Dallas, TX, USA), and the fission markers mitochondrial fission factor (MFF; Abnova; Cat# {"type":"entrez-protein","attrs":{"text":"PAB20518","term_id":"1236633578","term_text":"PAB20518"}} PAB20518 ) and dynamin-related protein (DRP)-1 (Cell Signaing; Cat# 8570).

Techniques: Double Immunofluorescence Staining, Marker, Fluorescence, Expressing