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: Molecular Neurobiology

Article Title: FTLD Patient–Derived Fibroblasts Show Defective Mitochondrial Function and Accumulation of p62

doi: 10.1007/s12035-021-02475-x

Figure Lengend Snippet: FTLD patient–derived fibroblasts show a trend of increased fission/fusion ratio, but no changes in phosphorylation of eIF2α. a Representative Western blot images from Fis1 and Mfn1+2 from fibroblast cell lysates. b Ratio of Fis1/ Mfn1+2. c Representative Western blot images from p-eIF2α and total eIF2α from fibroblast cell lysates. d Ratio of p-eIF2α/total eIF2α. Data are shown as the mean of three biological replicates ± SEM with ratio in controls set to 100. One-way ANOVA followed by Sidak’s multiple comparison test was performed. Only p values that were significant in the post hoc test are indicated in the graphs. n = 9 control, n = 9 FTLD with C9-HRE and n = 9 FTLD without C9-HRE

Article Snippet: The following primary antibodies were used: anti-Fission1 (1:1000, ALX-210-1037-0100; Enzo), anti-Mitofusin (1:1000, ab57602; Abcam), anti-eIF2α (1:1000, #9722; Cell Signaling Technology), anti-phospho-eIF2α (1:1000, #3597; Cell Signaling Technology), anti-pULK1Ser757 (1:1000, #14202S; Cell Signaling Technology), anti-ULK1 (1:1000, #8054; Cell Signaling Technology), anti-C9orf72 (1:500, 22637-1-AP; Proteintech), anti-SQSTM1/p62 (#5114, 1:1000; Cell Signaling Technology), anti-LC3B (1:3000, ab51520; Abcam), anti-poly-ubiquitinated proteins (FK1, 1:1000, BML-PW8805-0500; Enzo Life Sciences), anti-TDP-43 (1:1000, 10782-2-AP; Proteintech), anti-phospho-TDP-43 (1:1000, TIP-PTD-P02; CosmoBio), anti-beta-actin (1:1000, ab8226; Abcam), and anti-GAPDH (1:5000, ab8245; Abcam).

Techniques: Derivative Assay, Western Blot

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: Plant Physiology

Article Title: Peroxisome Function, Biogenesis, and Dynamics in Plants ^{1 [OPEN]}

doi: 10.1104/pp.17.01050

Figure Lengend Snippet: Peroxisome dynamics. Peroxisome biogenesis and division are coordinated by peroxins (numbered ovals) that coordinate peroxisomal membrane protein insertion into the ER or the peroxisomal membrane. After preperoxisomes bud from the ER, peroxisomes mature through import of matrix proteins. Peroxisomes can be degraded by pexophagy, a type of specialized autophagy. Dynamic peroxisome extensions (peroxules) assist peroxisome interactions with other organelles and can be associated with peroxisome division. PEX11 promotes peroxisome division together with a group of proteins (PMD1, FIS1, DRP) that also act in division of mitochondria or chloroplasts. PMP, peroxisomal membrane protein.

Article Snippet: The Arabidopsis paralogs of yeast FISSION1 ( Kemper et al., 2008 ), FIS1A and FIS1B, are tail-anchored membrane proteins acting in both mitochondrial and peroxisomal fission (for review, see Hu et al., 2012 ).

Techniques: Membrane