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: 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: Cell chemical biology

Article Title: The Mammalian Malonyl-CoA Synthetase ACSF3 Is Required for Mitochondrial Protein Malonylation and Metabolic Efficiency

doi: 10.1016/j.chembiol.2017.04.009

Figure Lengend Snippet: Key Resources Table

Article Snippet: Primary antibodies used include: ACSF3 rabbit polyclonal at 1:1000 (ThermoFisher PA5-25803), heat shock chaperone 70 (HSC70) mouse monoclonal at 1:1000 (Santa Cruz sc-7298), MitoProfile total OXPHOS mouse monoclonal antibody cocktail at 1:500 (Abcam ab110413), voltage-dependent anion channel 1 (VDAC1) rabbit polyclonal at 1:1000 (Calbiochem AB10527), aconitase 2 (ACO2) rabbit polyclonal at 1:1000 (Cell Signaling 6922), α-lipoic acid rabbit polyclonal at 1:1000 (Calbiochem #437695), phospho-ACC (Ser79) rabbit polyclonal at 1:1000 (Cell Signaling 11818), total ACC rabbit polyclonal at 1:1000 (Cell Signaling 3676), malonyl-lysine rabbit monoclonal mix at 1:1000 (Cell Signaling 14942), acetyl-lysine rabbit polyclonal at 1:1000 (Cell Signaling 9441), succinyl-lysine rabbit polyclonal at 1:1000 (PTM Biolabs 401), GFP mouse monoclonal at 1:1000 (Santa Cruz sc-9996).

Techniques: Magnetic Beads, Recombinant, Proliferation Assay, XF Assay, CyQUANT Assay, Activity Assay, Expressing, Plasmid Preparation, Transfection, Clone Assay