Journal: Stem Cells International

Article Title: Energy Metabolism and Lipidome Are Highly Regulated during Osteogenic Differentiation of Dental Follicle Cells

doi: 10.1155/2022/3674931

Figure Lengend Snippet: Expression of glycolysis and citric acid cycle markers during osteogenic differentiation of DFCs. (a) DFCs were cultured in osteogenic differentiation medium (ODM), BMP2 differentiation medium, or control medium (DMEM) for one, seven, 14, and 28 days before protein expression of the glycolysis markers hexokinase I, hexokinase II, phosphofructokinase, glycerinaldehyd-3-phosphat-dehydrogenase (GAPDH), pyruvate kinase M1/2, pyruvate kinase M2, and lactate dehydrogenase A, and protein expression of the citric acid cycle markers aconitase 2, isocitrate dehydrogenase 1, isocitrate dehydrogenase 2, dihydrolipoamide succinyltransferase (DLST), fumarase, citrase synthase, mitochondrial pyruvate carrier 1, and mitochondrial pyruvate carrier 2 were determined by western blot analysis. Quantification results and statistical analysis are shown in the Supplementary Table . (b–d) DFCs were cultured in osteogenic differentiation medium (ODM), BMP2 differentiation medium, or control medium (DMEM) for seven days before the amount of produced L-lactate (b) and hexokinase activity (c) were determined as markers of glycolysis and malate dehydrogenase activity was measured (d) as marker of citric acid cycle. Results are shown as means + standard deviation, and Student's t -test was performed to compare differentiation medium with control medium. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Subsequently, the membranes were incubated overnight at 4°C in a primary antibody against hypoxia-inducible factor 1-alpha (HIF-1 α ), cytochrome c, prohibitin 1, cytochrome c oxidase subunit 4 (COX IV), pyruvate dehydrogenase, succinate dehydrogenase complex subunit A (SDHA), heat shock protein 60 (HSP60), voltage-dependent anion channel (VDAC), hexokinase I, hexokinase II, phosphofructokinase, glycerinaldehyd-3-phosphat-dehydrogenase (GAPDH), pyruvate kinase M1/2, pyruvate kinase M2, lactate dehydrogenase A, aconitase 2, isocitrate dehydrogenase 1, isocitrate dehydrogenase 2, dihydrolipoamide succinyltransferase (DLST), fumarase, citrase synthase, mitochondrial pyruvate carrier 1, mitochondrial pyruvate carrier 2, acetyl-CoA carboxylase, phospho-acetyl-CoA carboxylase (Ser79), ATP-citrate lyase, phospho-ATP-citrate lyase (Ser455), acetyl-CoA synthetase, acyl-CoA synthetase, fatty acid synthase (all Cell Signaling Technology, Danvers, MA, USA), or elongation of very long chain fatty acids protein 6 (ELOVL6) (Abcam, Cambridge, UK).

Techniques: Expressing, Cell Culture, Western Blot, Produced, Activity Assay, Marker, Standard Deviation

Journal: Molecular Neurodegeneration

Article Title: Deficits in mitochondrial TCA cycle and OXPHOS precede rod photoreceptor degeneration during chronic HIF activation

doi: 10.1186/s13024-023-00602-x

Figure Lengend Snippet: Dysregulation of the citric acid cycle in rods with chronically active HIFs. a , b Significantly downregulated citric acid cycle associated enzymes in photoreceptors segments (PS) or outer nuclear layer (ONL) of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$rod^{\varDelta \ Vhl}$$\end{document} r o d Δ V h l mice at 2.5 months of age. Protein levels are shown as boxplots with the median and 25% and 75% percentiles ( a ) or bar graphs representing normalized abundance ratios of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$rod^{\varDelta \ Vhl}$$\end{document} r o d Δ V h l /ctrl ( b ). N = 6 mice per genotype. The samples with the highest (red) and lowest (blue) Cre levels are indicated. For statistical analysis on proteomics data see . nd, not detected. *: p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\le$$\end{document} ≤ 0.05. **: p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\le$$\end{document} ≤ 0.01. c Representation of the citric acid cycle with red arrows marking significantly downregulated proteins in the PRs of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$rod^{\varDelta \ Vhl}$$\end{document} r o d Δ V h l mice. d - f Representative immunofluorescence labeling for SDH subunit A (SDHA; d ), citrate synthase (CS; e ), and aconitate hydratase (ACO2; f ) in the PS regions of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$rod^{\varDelta \ Vhl}$$\end{document} r o d Δ V h l , \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$rod^{\varDelta \ Cox10}$$\end{document} r o d Δ C o x 10 and ctrl mice at time points as indicated (top panels). Pixel intensity (PI) profiles through the PS (bottom panels). White arrows: regions with reduced labeling. Scale bars, 50 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\upmu$$\end{document} μ m. N = 3 mice per genotype. g Dysregulation of OXPHOS and the TCA cycle in rods with chronic HIF activation is associated with impaired of OS biogenesis and leads to PRs degeneration

Article Snippet: The following primary antibodies were incubated overnight at 4 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$^{\circ }$$\end{document} ∘ C: rabbit anti-GLUT3 (1:250; Sigma-Aldrich, ab1344), mouse anti-COX4i1 (1:100; Abcam, ab14744), rabbit anti-CRE (1:300; Merck Millipore, 69050-3), rabbit anti-VDAC1 (1:250; Abcam, ab15895), rabbit anti-TFAM (1:250; Abcam, ab131607), rabbit anti-CS (1:250; GeneTex, GTX110624), rabbit anti-ACO2 (1:250; Cell Signaling Technology, 6571), rabbit anti-SDHA (1:250; Cell Signaling Technology, 11998) or rabbit anti-cARR (1:1000; Merck Millipore Chemicals, AB15282).

Techniques: Immunofluorescence, Labeling, Activation Assay

Journal: Journal of Cancer Prevention

Article Title: Breast Cancer Selective Disruption of Actin Cytoskeleton by Diallyl Trisulfide

doi: 10.15430/JCP.2022.27.2.101

Figure Lengend Snippet: (A) Reactome pathway enrichment analysis of downregulated genes by DATS treatment in MCF-10A and SK-BR-3 cell lines. (B) Immunoblot analysis for ACO2 and DLST in MCF-10A and SK-BR-3 cells treated with DMSO (control) or indicated doses of DATS for specified time points. The numbers above bands represent fold change in expression relative to corresponding DMSO-treated controls. (C) Quantification of ACO2 and DLST expression in SK-BR-3 cells. Results combined from three independent experiments are shown as mean ± SD (n = 3) and statistical analysis was done by one-way ANOVA followed by Dunnett’s test. DATS, diallyl trisulfide; DMSO, dimethyl sulfoxide, DLST, dihydrolipoamide S-succinyltransferase; ACO2, aconitase 2; HDR, Homology directed repair; ATR, ataxia telangiectasia and Rad3 related; AP, abasic sites; PCNA, proliferating cell nuclear antigen.

Article Snippet: Anti-aconitase 2 (ACO2) antibody and anti-dihydrolipoamide S-succinyltransferase (DLST) antibody were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Expressing

Journal: PLoS ONE

Article Title: The mitochondrial proteomic changes of rat hippocampus induced by 28-day simulated microgravity

doi: 10.1371/journal.pone.0265108

Figure Lengend Snippet: The protein changes of mitochondria of hippocampus after 28 days of tail suspension in rats.

Article Snippet: The primary antibodies used were aconitate hydratase (ACO2, 6571), dihydrolipoamide S-succinyltransferase (DLST, 11954), citrate synthase (CS, 14309), and VDAC (4661), and all of the antibodies were purchased from Cell Signaling Technology, Danvers, MA, USA.

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: The mitochondrial proteomic changes of rat hippocampus induced by 28-day simulated microgravity

doi: 10.1371/journal.pone.0265108

Figure Lengend Snippet: (A) Protein levels of ACO2, DLST and CS were determined by immunoblotting. (B) The relative expressions of ACO2, DLST and CS were represented by the intensity ratio between the protein and the loading control (VDAC) in each lane. n = 4, error bars indicated standard deviations. *p<0.05; **p<0.01 compared with each control group.

Article Snippet: The primary antibodies used were aconitate hydratase (ACO2, 6571), dihydrolipoamide S-succinyltransferase (DLST, 11954), citrate synthase (CS, 14309), and VDAC (4661), and all of the antibodies were purchased from Cell Signaling Technology, Danvers, MA, USA.

Techniques: Western Blot

Journal: iScience

Article Title: Prevention and regression of megamitochondria and steatosis by blocking mitochondrial fusion in the liver

doi: 10.1016/j.isci.2022.103996

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-ACO2 , Cell Signaling Technology , Cat# 6571; RRID: AB_2890191.

Techniques: Recombinant, Western Blot, Protease Inhibitor, Electron Microscopy, Activity Assay, Staining, Software