Journal: International journal of molecular sciences

Article Title: Role of Acyl-CoA Thioesterase 7 in Regulating Fatty Acid Metabolism and Its Contribution to the Onset and Progression of Bovine Clinical Mastitis.

doi: 10.3390/ijms252313046

Figure Lengend Snippet: Figure 4. Distribution analysis of ACOT7 in mammary glands of the two groups. (A) Pathological variation in bovine mammary glands of the healthy control group (Con/C; A1) and clinical mastitis groups (CM; A2) based on H&E staining. (B) ACOT7 distribution in bovine mammary glands of the Con/C (B1) and CM (B2) groups based on immunohistochemical staining. (C) Negative control (NC) for Con/C (C1) and CM (C2) groups. (D) Gray values of positive expression of ACOT7 protein quantified using ImageJ 1.44p software. Con/C, control group. CM, clinical mastitis group. NC, negative control; MA: mammary alveoli; MECs, mammary epithelial cells; NEUT, neutrophil. Scale bars, 50 µm (200× magnification). ** represents p < 0.01.

Article Snippet: The slices were incubated with a rabbit anti-ACOT7 primary antibody at a 1:200 dilution (Proteintech, Wuhan, China).

Techniques: Control, Staining, Immunohistochemical staining, Negative Control, Expressing, Software

Journal: International journal of molecular sciences

Article Title: Role of Acyl-CoA Thioesterase 7 in Regulating Fatty Acid Metabolism and Its Contribution to the Onset and Progression of Bovine Clinical Mastitis.

doi: 10.3390/ijms252313046

Figure Lengend Snippet: Figure 5. Co-localization analysis of ACOT7 in mammary glands of the two groups. (A–D) Co- localization analysis of ACOT7 in bovine mammary glands of the Con/C and CM groups: nuclei (blue, A1,A2), CK-18 (red, B1,B2), ACOT7 (green, C1,C2), merged with CK18 and ACOT7 (D1,D2) staining, respectively. (E) Positive IF signals of ACOT7 protein quantified using ImageJ software. Con/C, control group. CM, clinical mastitis group. MA, mammary alveoli; MECs, mammary epithelial cells; Scale bars, 50 µm (200× magnification). ** represents p < 0.01.

Article Snippet: The slices were incubated with a rabbit anti-ACOT7 primary antibody at a 1:200 dilution (Proteintech, Wuhan, China).

Techniques: Staining, Software, Control

Journal: International journal of molecular sciences

Article Title: Role of Acyl-CoA Thioesterase 7 in Regulating Fatty Acid Metabolism and Its Contribution to the Onset and Progression of Bovine Clinical Mastitis.

doi: 10.3390/ijms252313046

Figure Lengend Snippet: Figure 6. Expression patterns of ACOT7 mRNA and protein in mammary glands of the two groups. (A) Relative expression level of ACOT7 mRNA, monitored via qRT-PCR assays. (B) Protein bands and relative expression level of ACOT7, monitored via Western blot assays and using ImageJ software, respectively. Con/C, control group. CM, clinical mastitis group. Data are presented as means ± SEM. ** represents p < 0.01.

Article Snippet: The slices were incubated with a rabbit anti-ACOT7 primary antibody at a 1:200 dilution (Proteintech, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software, Control

Journal: International journal of molecular sciences

Article Title: Role of Acyl-CoA Thioesterase 7 in Regulating Fatty Acid Metabolism and Its Contribution to the Onset and Progression of Bovine Clinical Mastitis.

doi: 10.3390/ijms252313046

Figure Lengend Snippet: Figure 7. Proposed molecular mechanism underlying the effects of ACOT7 in the mammary glands of dairy cows with clinical mastitis (CM). MECs, mammary epithelial cells; FABP, fatty-acid-binding protein; FATP, fatty acid transport protein; TLR4, Toll-like receptor 4; TNF-α, tumor necrosis factor-α; IL-1β, interleukin-1β; LPS, lipopolysaccharide. Scale bar, 50 µm (200× magnification).

Article Snippet: The slices were incubated with a rabbit anti-ACOT7 primary antibody at a 1:200 dilution (Proteintech, Wuhan, China).

Techniques: Binding Assay

Journal: Nature Communications

Article Title: Convergent genomic signatures of flight loss in birds suggest a switch of main fuel

doi: 10.1038/s41467-019-10682-3

Figure Lengend Snippet: Identification of convergent evolutionary sites in flight-degenerate birds. a A phylogeny tree with American alligator as the outrgoup showing the convergence of eight studied flight-degenerate bird species. b Flying and flight-degenerate species were classified based on previous literatures and observations (Methods). Flight-degenerate species includes flightless birds and weak flyers. Average body mass of each species is plotted against its average wingspan measurement. c Two-dimensional frequency spectrum for nucleotides in flying and flight-degenerate species. The nucleotide frequencies of flight-degenerate birds were plotted against those of flying birds. The x -axis means the nucleotide counts among the eight flight-degenerate bird species and the y -axis means the nucleotide counts among 40 flying bird species. The number of loci is color-coded according to the scale on the right. d The pathway in which ATGL and ACOT7 are involved , . The main pathway for lipid metabolism is shadowed. In e and f , the upper shows the significant values (Fisher’s exact test) for the representative nucleotides on ATGL e and ACOT7 f , the middle shows the sequence of amino acids, and the bottom shows the schematic domains. The identified convergent sites are highlighted in red. Source data are provided as a Source Data file

Article Snippet: After the block with 5% skim milk, the membranes were incubated with primary antibodies (ACOT7 rabbit polyclonal antibody (1:1000, Proteintech, 15972-1-AP) and β-Actin mouse monoclonal antibody (internal control) (1:5000, Cell Signaling Technology, 8H10D10)) overnight at 4 °C followed by the incubation of secondary antibodies (goat anti-rabbit IgG-HRP (1:10,000, Santa Cruz Biotechnology, sc-2004) for ACOT7, donkey anti-mouse IgG-HRP (1:10,000, Abcam, ab97030) for β-Actin).

Techniques: Sequencing

Journal: Nature Communications

Article Title: Convergent genomic signatures of flight loss in birds suggest a switch of main fuel

doi: 10.1038/s41467-019-10682-3

Figure Lengend Snippet: Functional validation of ACOT7 GCG197GTG substitution. The abundances of free fatty acids were measured using GC–MS in the mouse adipocyte cells expressed with ACOT7 wild type ( fACOT7 ) and ACOT7 mutant type ( fACOT7mut ), respectively. For each free fatty acid, the abundance was compared between wild and mutant types and the median, first and third quartiles were shown in the box (middle bar represents median, upper bound the third quartile, lower bound the first quartile). Four independent experiments were performed. * P < 0.05 by the paired t test. Source data are provided as a Source Data file

Article Snippet: After the block with 5% skim milk, the membranes were incubated with primary antibodies (ACOT7 rabbit polyclonal antibody (1:1000, Proteintech, 15972-1-AP) and β-Actin mouse monoclonal antibody (internal control) (1:5000, Cell Signaling Technology, 8H10D10)) overnight at 4 °C followed by the incubation of secondary antibodies (goat anti-rabbit IgG-HRP (1:10,000, Santa Cruz Biotechnology, sc-2004) for ACOT7, donkey anti-mouse IgG-HRP (1:10,000, Abcam, ab97030) for β-Actin).

Techniques: Functional Assay, Biomarker Discovery, Gas Chromatography-Mass Spectrometry, Mutagenesis

Journal: Nature Communications

Article Title: Convergent genomic signatures of flight loss in birds suggest a switch of main fuel

doi: 10.1038/s41467-019-10682-3

Figure Lengend Snippet: Modeling simulations of effects of functional changes on the energy landscape. The actual abundance of acyl-CoA a and acetyl-CoA b was measured in three flight-degenerate (turkey, chicken and ostrich) and three flying species (zebra finch, parrot and pigeon) using LC–MS. The simulated results for ATGL and ACOT7 are shown in c and d respectively. * P < 0.05, *** P < 0.001 by Wilcoxon rank sum test. Error bars indicate s.e.m. Source data are provided as a Source Data file

Article Snippet: After the block with 5% skim milk, the membranes were incubated with primary antibodies (ACOT7 rabbit polyclonal antibody (1:1000, Proteintech, 15972-1-AP) and β-Actin mouse monoclonal antibody (internal control) (1:5000, Cell Signaling Technology, 8H10D10)) overnight at 4 °C followed by the incubation of secondary antibodies (goat anti-rabbit IgG-HRP (1:10,000, Santa Cruz Biotechnology, sc-2004) for ACOT7, donkey anti-mouse IgG-HRP (1:10,000, Abcam, ab97030) for β-Actin).

Techniques: Functional Assay, Liquid Chromatography with Mass Spectroscopy

Journal: Nature Communications

Article Title: Convergent genomic signatures of flight loss in birds suggest a switch of main fuel

doi: 10.1038/s41467-019-10682-3

Figure Lengend Snippet: Ancestral state reconstruction for ATGL 321 and ACOT7 197 codons. The avian phylogeny from the previous study that we contributed to was used with the American alligator genome used as the outgroup

Article Snippet: After the block with 5% skim milk, the membranes were incubated with primary antibodies (ACOT7 rabbit polyclonal antibody (1:1000, Proteintech, 15972-1-AP) and β-Actin mouse monoclonal antibody (internal control) (1:5000, Cell Signaling Technology, 8H10D10)) overnight at 4 °C followed by the incubation of secondary antibodies (goat anti-rabbit IgG-HRP (1:10,000, Santa Cruz Biotechnology, sc-2004) for ACOT7, donkey anti-mouse IgG-HRP (1:10,000, Abcam, ab97030) for β-Actin).

Techniques:

Journal: Nature communications

Article Title: Convergent genomic signatures of flight loss in birds suggest a switch of main fuel.

doi: 10.1038/s41467-019-10682-3

Figure Lengend Snippet: Fig. 1 Identification of convergent evolutionary sites in flight-degenerate birds. a A phylogeny tree5 with American alligator as the outrgoup showing the convergence of eight studied flight-degenerate bird species. b Flying and flight-degenerate species were classified based on previous literatures and observations (Methods). Flight-degenerate species includes flightless birds and weak flyers. Average body mass of each species is plotted against its average wingspan measurement. c Two-dimensional frequency spectrum for nucleotides in flying and flight-degenerate species. The nucleotide frequencies of flight-degenerate birds were plotted against those of flying birds. The x-axis means the nucleotide counts among the eight flight-degenerate bird species and the y-axis means the nucleotide counts among 40 flying bird species. The number of loci is color-coded according to the scale on the right. d The pathway in which ATGL and ACOT7 are involved35,70. The main pathway for lipid metabolism is shadowed. In e and f, the upper shows the significant values (Fisher’s exact test) for the representative nucleotides on ATGL e and ACOT7 f, the middle shows the sequence of amino acids, and the bottom shows the schematic domains. The identified convergent sites are highlighted in red. Source data are provided as a Source Data file

Article Snippet: After the block with 5% skim milk, the membranes were incubated with primary antibodies (ACOT7 rabbit polyclonal antibody (1:1000, Proteintech, 15972-1-AP) and β-Actin mouse monoclonal antibody (internal control) (1:5000, Cell Signaling Technology, 8H10D10)) overnight at 4 °C followed by the incubation of secondary antibodies (goat antirabbit IgG-HRP (1:10,000, Santa Cruz Biotechnology, sc-2004) for ACOT7, donkey anti-mouse IgG-HRP (1:10,000, Abcam, ab97030) for β-Actin).

Techniques: Sequencing

Journal: Nature communications

Article Title: Convergent genomic signatures of flight loss in birds suggest a switch of main fuel.

doi: 10.1038/s41467-019-10682-3

Figure Lengend Snippet: Fig. 4 Modeling simulations of effects of functional changes on the energy landscape. The actual abundance of acyl-CoA a and acetyl-CoA b was measured in three flight-degenerate (turkey, chicken and ostrich) and three flying species (zebra finch, parrot and pigeon) using LC–MS. The simulated results for ATGL and ACOT7 are shown in c and d respectively. *P < 0.05, ***P < 0.001 by Wilcoxon rank sum test. Error bars indicate s.e.m. Source data are provided as a Source Data file

Article Snippet: After the block with 5% skim milk, the membranes were incubated with primary antibodies (ACOT7 rabbit polyclonal antibody (1:1000, Proteintech, 15972-1-AP) and β-Actin mouse monoclonal antibody (internal control) (1:5000, Cell Signaling Technology, 8H10D10)) overnight at 4 °C followed by the incubation of secondary antibodies (goat antirabbit IgG-HRP (1:10,000, Santa Cruz Biotechnology, sc-2004) for ACOT7, donkey anti-mouse IgG-HRP (1:10,000, Abcam, ab97030) for β-Actin).

Techniques: Functional Assay, Liquid Chromatography with Mass Spectroscopy

Journal: Scientific Reports

Article Title: Increasing Acyl CoA thioesterase activity alters phospholipid profile without effect on insulin action in skeletal muscle of rats

doi: 10.1038/s41598-018-32354-w

Figure Lengend Snippet: Overexpression of AAV9-tMCK-Acot7 and its effect on the opposing ACSL enzyme in tibialis muscle. Acot7 protein levels ( A ), thioesterase activity ( B ) ACSL1 protein level ( C ) Acyl-CoA synthase activity ( D ). Data is mean ± SEM, *p < 0.05, 2-way ANOVA, diet; † p < 0.01 2-way ANOVA, Acot7; # p < 0.01 post-hoc Holm and Sidak test, (n = 6).

Article Snippet: Protein quantification was carried out by immunoblotting using 1:1000 dilutions of antibodies for Acot7 (Abcam85151), ACSL1 (Cell Signalling 4047), Mitochondrial complex (Mitosciences601), VDAC (Cell SignalingS4866), UCP3 (Abcam4866) as described .

Techniques: Over Expression, Activity Assay