Journal: PLoS ONE

Article Title: AdipoRon, adiponectin receptor agonist, improves vascular function in the mesenteric arteries of type 2 diabetic mice

doi: 10.1371/journal.pone.0230227

Figure Lengend Snippet: (A) CTR: control mice treated with vehicle, DB: diabetic mice treated with vehicle, and DB+ADI (AdipoRon): diabetic mice treated with AdipoRon. Representative western blot analysis and quantitative data for MYPT1 in mesenteric arteires (n = 4). (B) Vehicle: VSMCs stimulated with vehicle, PE: VSMCs stimulated with phenylephrine (5 μM), PE+ADI: VSMCs co-stimulated with PE (5 μM) and AdipoRon (ADI, 50 μM). Representative western blot analysis and quantitative data for MYPT1 in VSMCs (n = 4). Each group of blots was cropped from a same gel and membrane. * P < 0.05.

Article Snippet: Western blot analysis was performed for AdiR1 (1:1000 dilution; Abcam #ab70362, Cambridge, MA, USA), AdiR2 (1:1000 dilution; Abcam #ab77613, Cambridge, MA, USA), APPL1 (1:1000 dilution; Santa Cruz Biotechnology #SC-271901, Santa Cruz, CA, USA), APPL2 (1:1000 dilution; Santa Cruz Biotechnology #SC-271084, Santa Cruz, CA, USA), phosphorylated AMPK (1:1000 dilution; Abcam #2532s, Cambridge, MA, USA), total AMPK (1:1000 dilution; Abcam #2535s, Cambridge, MA, USA), phosphorylated eNOS (1:1000 dilution; Cell signaling #9571, Danvers, MA, USA), total eNOS (1:1000 dilution; Cell signaling #9586s, Danvers, MA, USA), and phosphorylated MYPT1 (myosin phosphatase target subunit 1, 1:500 dilution; Bioss #bs-3288R, Woburn, MA, USA), and total MYPT1 (1:500 dilution; Bioss #bs-3787R, Woburn, MA, USA).

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Inhibitory effects of fenretinide metabolites N-[4-methoxyphenyl]retinamide (MPR) and 4-oxo-N-(4-hydroxyphenyl)retinamide (3-keto-HPR) on fenretinide molecular targets β-carotene oxygenase 1, stearoyl-CoA desaturase 1 and dihydroceramide Δ4-desaturase 1

doi: 10.1371/journal.pone.0176487

Figure Lengend Snippet: A) SCD1 protein expression is decreased after treatment with fenretinide, and its metabolite 3-keto-HPR. ARPE-19 cells were treated with the indicated concentration of fenretinide, its metabolites, A1120, or vehicle for 24 h. Cell lysates were analyzed by Western blotting using mouse monoclonal antibody against SCD1 (upper panel) in comparison with immunoreactivity to α-tubulin which was employed as a loading control (lower panel) and densitometry for each lane was performed and analyzed by ImageJ to provide the SCD1/α-tubulin ratio for each lane. B) SCD1 protein activity is decreased after treatment with fenretinide, and its metabolite 3-keto-HPR. ARPE-19 cells were treated with the indicated concentration of fenretinide, its metabolites, A1120, or vehicle for 16 h, then incubated an additional 5 h in the presence of methyl D3-palmitic acid (50 μM) before harvesting. Free fatty acids present in lipid extracts were analyzed by LC/MS and SCD1 activity, measured by the conversion of methyl-D3 palmitic acid to methyl-D3 palmitoleic acid, was determined. Fenretinide and 3-keto-HPR are effective inhibitors of SCD1 activity, whereas MPR and A1120 have no inhibitory effect on SCD1 activity at all. Lipids were extracted from two 100 mm Petri dishes and combined for mass spectrometry analysis. Data plotted as means ± standard deviation; n = 3.

Article Snippet: After blocking in 5% non-fat milk in Tris-buffered saline (TBS) containing 0.01% Tween 20 for 1 h, the membranes were incubated overnight at 4°C with mouse monoclonal anti-SCD1 antibody (1:200; Kamiya Biomedical Company).

Techniques: Expressing, Concentration Assay, Western Blot, Activity Assay, Incubation, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Standard Deviation

Journal: Biology of Reproduction

Article Title: Stearoyl-CoA desaturase activity in bovine cumulus cells protects the oocyte against saturated fatty acid stress

doi: 10.1095/biolreprod.116.146159

Figure Lengend Snippet: Details of primers used for PCR.

Article Snippet: SCD1 , Stearoyl-CoA-desaturase 1 , Custom , Rabbit , Bovine , Pacific Immunology , 1:1000.

Techniques: Sequencing

Journal: Biology of Reproduction

Article Title: Stearoyl-CoA desaturase activity in bovine cumulus cells protects the oocyte against saturated fatty acid stress

doi: 10.1095/biolreprod.116.146159

Figure Lengend Snippet: Details of antibodies used for western blotting.

Article Snippet: SCD1 , Stearoyl-CoA-desaturase 1 , Custom , Rabbit , Bovine , Pacific Immunology , 1:1000.

Techniques: Western Blot

Journal: Biology of Reproduction

Article Title: Stearoyl-CoA desaturase activity in bovine cumulus cells protects the oocyte against saturated fatty acid stress

doi: 10.1095/biolreprod.116.146159

Figure Lengend Snippet: SCD1 gene and protein expression in cumulus cells and oocytes. (A) The increase of SCD1 mRNA expression in cumulus cells of COCs at 23 h of maturation in comparison to 0 h of maturation, maturation in control medium (green bar), and after exposure to stearic and palmitic acid (100 and 150 μM; purple bar), and stearic, palmitic, and oleic acid (100, 150 and 200 μM; blue bar). Results are presented as mean ± SD; data with different letters differ significantly ( P ≤ 0.01). (B) Representative immunoblot showing expression of SCD1 and actin in oocyte and cumulus cells. SCD1 protein expression in oocytes and cumulus cells is shown at 0 h and after 23 h of maturation in control medium and in the presence of 250 μM stearic acid.

Article Snippet: SCD1 , Stearoyl-CoA-desaturase 1 , Custom , Rabbit , Bovine , Pacific Immunology , 1:1000.

Techniques: Expressing, Comparison, Control, Western Blot

Journal: Biology of Reproduction

Article Title: Stearoyl-CoA desaturase activity in bovine cumulus cells protects the oocyte against saturated fatty acid stress

doi: 10.1095/biolreprod.116.146159

Figure Lengend Snippet: SCD activity in cumulus cells prevents negative impact of saturated free fatty acid on oocyte developmental competence. Percentages of cleavage (A, C, E) and blastocyst formation (B, D, F) of oocytes from COCs matured in control medium (green bars) and in the presence of (A, B) saturated stearic acid (250 μM; orange bar), (C, D) palmitic acid (250 μM; red bar), or (E and F) saturated stearic and palmitic acid (100 and 150 μM; purple bar) and with SCD inhibition (white bars with outline in color of experimental condition). The SCD1 inhibitors were from Biovision (BIO) or Cayman (CAY). Results are presented as mean ± SD; data with different letters differ significantly ( P < 0.05).

Article Snippet: SCD1 , Stearoyl-CoA-desaturase 1 , Custom , Rabbit , Bovine , Pacific Immunology , 1:1000.

Techniques: Activity Assay, Control, Inhibition

Journal: Biology of Reproduction

Article Title: Stearoyl-CoA desaturase activity in bovine cumulus cells protects the oocyte against saturated fatty acid stress

doi: 10.1095/biolreprod.116.146159

Figure Lengend Snippet: Apoptosis and necrosis rates in COCs after exposure to stearic acid with and without SCD1 activity. SCD1 activity was inhibited (white bars with outline in the color of experimental condition) in the control group (green bars) and in the presence of saturated stearic acid (250 μM; orange bar). In panel A, the apoptosis levels in cumulus cells from the different conditions are presented; panel B presents the necrosis levels. Results are presented as mean ± SD. Data with different letters differ significantly ( P < 0.05). BIO = SCD1 inhibitor from Biovision.

Article Snippet: SCD1 , Stearoyl-CoA-desaturase 1 , Custom , Rabbit , Bovine , Pacific Immunology , 1:1000.

Techniques: Activity Assay, Control

Journal: Biology of Reproduction

Article Title: Stearoyl-CoA desaturase activity in bovine cumulus cells protects the oocyte against saturated fatty acid stress

doi: 10.1095/biolreprod.116.146159

Figure Lengend Snippet: The molar ratio of C18:1/C18:0 based on the levels in matured cumulus cells. SCD1 activity was inhibited (white bars with outline in the color of experimental condition) in the control group (green bars) and in the presence of saturated stearic acid (250 μM; orange bars) during maturation. Panel A presents the C18:1/C18:0 ratio based on the levels of C18:0 presented in panel B and C18:1 in panel C in the lipid of cumulus cells from matured COCs. Results are presented as mean ± SD. Data with different letters differ significantly ( P < 0.05). BIO = SCD1 inhibitor from Biovision. CPS = ion counts per second.

Article Snippet: SCD1 , Stearoyl-CoA-desaturase 1 , Custom , Rabbit , Bovine , Pacific Immunology , 1:1000.

Techniques: Activity Assay, Control

Journal: Biology of Reproduction

Article Title: Stearoyl-CoA desaturase activity in bovine cumulus cells protects the oocyte against saturated fatty acid stress

doi: 10.1095/biolreprod.116.146159

Figure Lengend Snippet: Lipid droplet formation in cumulus cells after exposure to stearic and oleic acid without and with SCD1 inhibition. Panel A shows the confocal images of lipid droplets (green) and cumulus cell nuclei (red) for the different maturation groups; control, stearic, and oleic acid without and with a SCD1 inhibitor; (B) Graph representing the number of lipid droplets in cumulus cells after maturation in control medium (green bar), and in the presence of stearic acid (orange bar) or oleic acid (blue bar) during 23 h of maturation without and with inhibition of SCD1 activity (white bars with outline in the color of experimental condition); (C) Graph representing the mean size of lipid droplets, data are normalized on the basis of the control group without SCD1 inhibition. The average diameter of lipid droplets in the control group was 0.99 ± 0.14 μm. Results are presented as mean ± SD. Data with different letters differ significantly ( P < 0.05). BIO = SCD1 inhibitor from Biovision. Scale bar represents 20 μm.

Article Snippet: SCD1 , Stearoyl-CoA-desaturase 1 , Custom , Rabbit , Bovine , Pacific Immunology , 1:1000.

Techniques: Inhibition, Control, Activity Assay