Journal: Cells

Article Title: Therapeutic Potential of Stearoyl-CoA Desaturase1 (SCD1) in Modulating the Effects of Fatty Acids on Osteoporosis

doi: 10.3390/cells13211781

Figure Lengend Snippet: Schematic diagram illustrating the research concept. In osteoporosis models, fatty acids such as SFA and MUFA are secreted from expanding MAT. This study suggests that SCD1 can promote bone regeneration by converting SFA, which is known to have a negative effect on osteoblast differentiation and MSC proliferation, into MUFA, which can have a positive effect on bone metabolism. This highlights the therapeutic potential of MSCs through SCD1 overexpression in bone regeneration in osteoporosis models.

Article Snippet: After blocking with 5% bovine serum albumin (BSA) in TBST (Tris-buffered saline with Tween 20) for 1 h, the membranes were washed and incubated sequentially with a primary anti-SCD1 antibody (#2794, Cell Signaling Technology, Beverly, MA, USA) and anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibodies.

Techniques: Over Expression

Journal: Cells

Article Title: Therapeutic Potential of Stearoyl-CoA Desaturase1 (SCD1) in Modulating the Effects of Fatty Acids on Osteoporosis

doi: 10.3390/cells13211781

Figure Lengend Snippet: Effect of SCD1 overexpression on osteogenic differentiation reduced by palmitic acid (PA). ( A ) Production of adenovirus-based SCD1 vector. ( B ) Confirmation of cell viability of BM-MSCs according to virus MOI. ( C ) Verification of MOI-dependent overexpression of Ad-SCD1. ( D ) Confirmation of normal virus infection and GFP expression in BM-MSCs. ( E ) Measurement of cell viability with the virus. ( F – I ) Confirmation of the effect of Ad-SCD1 on osteogenic differentiation of PA-treated BM-MSCs through comparison of ALP and ARS staining, ALP activity, and calcium deposition. (*** p < 0.001).

Article Snippet: After blocking with 5% bovine serum albumin (BSA) in TBST (Tris-buffered saline with Tween 20) for 1 h, the membranes were washed and incubated sequentially with a primary anti-SCD1 antibody (#2794, Cell Signaling Technology, Beverly, MA, USA) and anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibodies.

Techniques: Over Expression, Plasmid Preparation, Virus, Infection, Expressing, Comparison, Staining, Activity Assay

Journal: Cells

Article Title: Therapeutic Potential of Stearoyl-CoA Desaturase1 (SCD1) in Modulating the Effects of Fatty Acids on Osteoporosis

doi: 10.3390/cells13211781

Figure Lengend Snippet: In vivo experiments to verify the effect of SCD1 on bone defect recovery in an osteoporosis model. ( A ) Schematic diagram of the animal modeling experiment. ( B ) X-ray scan images of the left distal femur at 1, 3, 5, and 7 weeks after cell transplantation into the defect. ( C ) BMD measurements by DEXA imaging at 1, 3, 5, and 7 weeks (* p < 0.05, *** p < 0.001).

Article Snippet: After blocking with 5% bovine serum albumin (BSA) in TBST (Tris-buffered saline with Tween 20) for 1 h, the membranes were washed and incubated sequentially with a primary anti-SCD1 antibody (#2794, Cell Signaling Technology, Beverly, MA, USA) and anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibodies.

Techniques: In Vivo, Transplantation Assay, Imaging

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: Figure 2. 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: The membranes were rinsed in TBS-Tween [(TBS with 0.05% Tween-20 (ICN, Aurora OH, USA)] and blocked with 5% nonfat dry milk in TBS-Tween (blocking buffer) for 1 h and probed with primary SCD1 antibody (custom rabbit anti-bovine SCD1, Pacific Immunology, Ramona CA, USA; a kind gift of Dr. Benjamin Corl, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA) or with a polyclonal antibody against ß-actin (sc-1616, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:1000 in blocking buffer at 4◦C overnight.

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: Figure 3. 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: The membranes were rinsed in TBS-Tween [(TBS with 0.05% Tween-20 (ICN, Aurora OH, USA)] and blocked with 5% nonfat dry milk in TBS-Tween (blocking buffer) for 1 h and probed with primary SCD1 antibody (custom rabbit anti-bovine SCD1, Pacific Immunology, Ramona CA, USA; a kind gift of Dr. Benjamin Corl, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA) or with a polyclonal antibody against ß-actin (sc-1616, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:1000 in blocking buffer at 4◦C overnight.

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: Figure 4. 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: The membranes were rinsed in TBS-Tween [(TBS with 0.05% Tween-20 (ICN, Aurora OH, USA)] and blocked with 5% nonfat dry milk in TBS-Tween (blocking buffer) for 1 h and probed with primary SCD1 antibody (custom rabbit anti-bovine SCD1, Pacific Immunology, Ramona CA, USA; a kind gift of Dr. Benjamin Corl, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA) or with a polyclonal antibody against ß-actin (sc-1616, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:1000 in blocking buffer at 4◦C overnight.

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: Figure 5. 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: The membranes were rinsed in TBS-Tween [(TBS with 0.05% Tween-20 (ICN, Aurora OH, USA)] and blocked with 5% nonfat dry milk in TBS-Tween (blocking buffer) for 1 h and probed with primary SCD1 antibody (custom rabbit anti-bovine SCD1, Pacific Immunology, Ramona CA, USA; a kind gift of Dr. Benjamin Corl, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA) or with a polyclonal antibody against ß-actin (sc-1616, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:1000 in blocking buffer at 4◦C overnight.

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: Figure 6. 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: The membranes were rinsed in TBS-Tween [(TBS with 0.05% Tween-20 (ICN, Aurora OH, USA)] and blocked with 5% nonfat dry milk in TBS-Tween (blocking buffer) for 1 h and probed with primary SCD1 antibody (custom rabbit anti-bovine SCD1, Pacific Immunology, Ramona CA, USA; a kind gift of Dr. Benjamin Corl, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA) or with a polyclonal antibody against ß-actin (sc-1616, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:1000 in blocking buffer at 4◦C overnight.

Techniques: Inhibition, Control, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: α-Tocopherol-13′-Carboxychromanol Induces Cell Cycle Arrest and Cell Death by Inhibiting the SREBP1-SCD1 Axis and Causing Imbalance in Lipid Desaturation

doi: 10.3390/ijms24119229

Figure Lengend Snippet: α-T-13′-COOH suppresses proliferation of murine macrophages. The influence of α-T-13′-COOH and the SCD1 inhibitor CAY10566 on the proliferation of murine RAW264.7 cells was analyzed by colorimetric detection of BrdU incorporation into DNA synthesized during cell division. Cells were treated with either vehicle control (DMSO, w/o), α-T-13′-COOH (0.5 µM, 5 µM), the SCD1 inhibitor CAY10566 (100 nM) in serum-free medium, or FBS (10% ( v / v )) for 24 h. Experiments with α-T-13′-COOH and CAY10566 were carried out in independent experiments. Fold changes (FCs) relative to the respective vehicle-treated control (w/o) are shown. Points represent data from six or seven independent biological replicates and their means ± standard error of the mean (SEM) are shown. p -Values were calculated using repeated measurement one-way ANOVA with Tukey’s post-hoc test. *, p < 0.05, **, p < 0.01.

Article Snippet: Primary antibodies were directed against murine Scd1 (1:1000; rabbit anti-Scd1; #2438, Cell Signaling, Frankfurt am Main, Germany), Fasn (1:1000, rabbit anti-Fasn; #3189, Cell Signaling), Srebp1 (1:200 to 1:400, mouse anti-Srebp1; sc-13551, Santa Cruz Biotechnology, Heidelberg, Germany), Hdac1 (1:1000, mouse anti-Hdac1; sc-81598, Santa Cruz Biotechnology), β-actin (1:1000, mouse anti-β-actin; #3700, Cell Signaling), and α-tubulin (1:5000, mouse anti-α-tubulin; clone B-5-1-2, BD Biosciences, Heidelberg, Germany).

Techniques: BrdU Incorporation Assay, Synthesized, Control

Journal: International Journal of Molecular Sciences

Article Title: α-Tocopherol-13′-Carboxychromanol Induces Cell Cycle Arrest and Cell Death by Inhibiting the SREBP1-SCD1 Axis and Causing Imbalance in Lipid Desaturation

doi: 10.3390/ijms24119229

Figure Lengend Snippet: α-T-13′-COOH augments the proportion of cells in the G 1 /G 0 phase. Cell cycle analysis ( A ) and quantification of each cell cycle phase ( B ) were performed by flow cytometry following propidium iodide staining. RAW264.7 cells were incubated with either vehicle (DMSO, ‘w/o’), α-T-13′-COOH (0.5 µM, 5.0 µM), the SCD1 inhibitor CAY10566 (100 nM), or FBS (10% ( v / v )), which served as positive control. Studies with α-T-13′-COOH and CAY10566 were carried out in independent measurements. Data from five independent biological replicates are presented as means − SEM. Statistical significance was evaluated using repeated measurement two-way ANOVA with Tukey’s post-hoc test. ***, p < 0.001 (data of G 1 /G 0 phase vs. w/o); &&&, p < 0.001 (data of S phase vs. w/o); $, p < 0.05; $$, p < 0.01 (data of G 2 /M phase vs. w/o).

Article Snippet: Primary antibodies were directed against murine Scd1 (1:1000; rabbit anti-Scd1; #2438, Cell Signaling, Frankfurt am Main, Germany), Fasn (1:1000, rabbit anti-Fasn; #3189, Cell Signaling), Srebp1 (1:200 to 1:400, mouse anti-Srebp1; sc-13551, Santa Cruz Biotechnology, Heidelberg, Germany), Hdac1 (1:1000, mouse anti-Hdac1; sc-81598, Santa Cruz Biotechnology), β-actin (1:1000, mouse anti-β-actin; #3700, Cell Signaling), and α-tubulin (1:5000, mouse anti-α-tubulin; clone B-5-1-2, BD Biosciences, Heidelberg, Germany).

Techniques: Cell Cycle Assay, Flow Cytometry, Staining, Incubation, Positive Control

Journal: International Journal of Molecular Sciences

Article Title: α-Tocopherol-13′-Carboxychromanol Induces Cell Cycle Arrest and Cell Death by Inhibiting the SREBP1-SCD1 Axis and Causing Imbalance in Lipid Desaturation

doi: 10.3390/ijms24119229

Figure Lengend Snippet: α-T-13′-COOH induces apoptosis in murine macrophages. Representative flow cytometry plots ( A ) and quantification of early apoptosis ( B ), late apoptosis, and necrosis ( C ) by staining with 7-AAD and PE-labelled Annexin V. RAW264.7 cells were cultivated with either vehicle (DMSO, ‘w/o’), α-T-13′-COOH (0.5 µM, 5 µM), the SCD1 inhibitor CAY10566 (100 nM), or the positive control camptothecin (CPT, 2.5 µM) for 24 h. Experiments with α-T-13′-COOH and CAY10566 were performed independently from each other. Relative changes in the number of apoptotic or necrotic cells compared to the vehicle control are shown. Means ± SEM of four or five independent experiments (dots) are shown. p -Values were calculated using repeated measurement one-way ANOVA with Tukey’s post hoc test. *, p < 0.05, **, p < 0.01; ***, p < 0.001 compared with w/o.

Article Snippet: Primary antibodies were directed against murine Scd1 (1:1000; rabbit anti-Scd1; #2438, Cell Signaling, Frankfurt am Main, Germany), Fasn (1:1000, rabbit anti-Fasn; #3189, Cell Signaling), Srebp1 (1:200 to 1:400, mouse anti-Srebp1; sc-13551, Santa Cruz Biotechnology, Heidelberg, Germany), Hdac1 (1:1000, mouse anti-Hdac1; sc-81598, Santa Cruz Biotechnology), β-actin (1:1000, mouse anti-β-actin; #3700, Cell Signaling), and α-tubulin (1:5000, mouse anti-α-tubulin; clone B-5-1-2, BD Biosciences, Heidelberg, Germany).

Techniques: Flow Cytometry, Staining, Positive Control, Control

Journal: International Journal of Molecular Sciences

Article Title: α-Tocopherol-13′-Carboxychromanol Induces Cell Cycle Arrest and Cell Death by Inhibiting the SREBP1-SCD1 Axis and Causing Imbalance in Lipid Desaturation

doi: 10.3390/ijms24119229

Figure Lengend Snippet: α-T-13′-COOH reduces the estimated enzyme activity of SCD1. The desaturation activity of SCD1 is estimated as the product-to-substrate ratios (C16:1/C16:0 and C18:1/C18:0) of lipid species (( A ), TGs; ( B ), PC). Experiments were performed in four independent biological replicates (dots). Data are presented as means ± SEM. p -Values were calculated using repeated measurement one-way ANOVA with Tukey’s post-hoc test. **, p < 0.01; ***, p < 0.001; (w/o vs. α-T-13′-COOH).

Article Snippet: Primary antibodies were directed against murine Scd1 (1:1000; rabbit anti-Scd1; #2438, Cell Signaling, Frankfurt am Main, Germany), Fasn (1:1000, rabbit anti-Fasn; #3189, Cell Signaling), Srebp1 (1:200 to 1:400, mouse anti-Srebp1; sc-13551, Santa Cruz Biotechnology, Heidelberg, Germany), Hdac1 (1:1000, mouse anti-Hdac1; sc-81598, Santa Cruz Biotechnology), β-actin (1:1000, mouse anti-β-actin; #3700, Cell Signaling), and α-tubulin (1:5000, mouse anti-α-tubulin; clone B-5-1-2, BD Biosciences, Heidelberg, Germany).

Techniques: Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: α-Tocopherol-13′-Carboxychromanol Induces Cell Cycle Arrest and Cell Death by Inhibiting the SREBP1-SCD1 Axis and Causing Imbalance in Lipid Desaturation

doi: 10.3390/ijms24119229

Figure Lengend Snippet: mRNA expression of genes involved in de novo lipogenesis of MUFAs is reduced by α-T-13′-COOH in murine RAW264.7 macrophages. The impact of α-T-13′-COOH on mRNA expression of Fasn, Scd1, Scd2, Acc1, and Acc2 was determined by RT-qPCR under serum-free conditions. RAW264.7 cells were incubated with either vehicle (DMSO, ‘w/o’) or 0.5 or 5.0 µM α-T-13′-COOH for the indicated times (6 or 24 h). Expression levels of target genes were normalized to the expression of the reference gene Ppib which was not affected. FCs were calculated vs. the respective vehicle-treated control and the log 2 (FC) is shown in the figure. Experiments were performed in five independent biological replicates (dots). The data are also shown as part of the heatmap in . Data are presented as means ± SEM. Statistical comparisons were made by repeated measurement two-way ANOVA with Bonferroni’s post-hoc test between control and α-T-13′-COOH-treated samples. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: Primary antibodies were directed against murine Scd1 (1:1000; rabbit anti-Scd1; #2438, Cell Signaling, Frankfurt am Main, Germany), Fasn (1:1000, rabbit anti-Fasn; #3189, Cell Signaling), Srebp1 (1:200 to 1:400, mouse anti-Srebp1; sc-13551, Santa Cruz Biotechnology, Heidelberg, Germany), Hdac1 (1:1000, mouse anti-Hdac1; sc-81598, Santa Cruz Biotechnology), β-actin (1:1000, mouse anti-β-actin; #3700, Cell Signaling), and α-tubulin (1:5000, mouse anti-α-tubulin; clone B-5-1-2, BD Biosciences, Heidelberg, Germany).

Techniques: Expressing, Quantitative RT-PCR, Incubation, Control

Journal: International Journal of Molecular Sciences

Article Title: α-Tocopherol-13′-Carboxychromanol Induces Cell Cycle Arrest and Cell Death by Inhibiting the SREBP1-SCD1 Axis and Causing Imbalance in Lipid Desaturation

doi: 10.3390/ijms24119229

Figure Lengend Snippet: The amount of Scd1 protein is reduced by α-T-13′-COOH. The expression of Scd1 protein (≈37 kDa) was investigated by Western blotting of whole cell lysates. RAW264.7 macrophages were treated with either vehicle (DMSO, ‘w/o’) or 0.5 or 5.0 µM α-T-13′-COOH for 8 or 24 h. The signals of Scd1 and Fasn were normalized to the signal of the loading control β-actin (≈45 kDa) or α-tubulin (≈55 kDa), respectively. The graphs show the fold changes of Scd1 protein levels ( A ) or Fasn protein levels ( B ) in cells treated with α-T-13′-COOH relative to vehicle-treated control cells (w/o). Representative immunoblots are shown in the panels below each graph. Experiments were performed in three independent biological replicates (dots). Unprocessed Western blot images are shown in . Data are presented as means ± SEM. Statistical comparisons were made by repeated measurement two-way ANOVA with Tukey’s post-hoc test between control and α-T-13′-COOH-treated samples. ***, p < 0.001.

Article Snippet: Primary antibodies were directed against murine Scd1 (1:1000; rabbit anti-Scd1; #2438, Cell Signaling, Frankfurt am Main, Germany), Fasn (1:1000, rabbit anti-Fasn; #3189, Cell Signaling), Srebp1 (1:200 to 1:400, mouse anti-Srebp1; sc-13551, Santa Cruz Biotechnology, Heidelberg, Germany), Hdac1 (1:1000, mouse anti-Hdac1; sc-81598, Santa Cruz Biotechnology), β-actin (1:1000, mouse anti-β-actin; #3700, Cell Signaling), and α-tubulin (1:5000, mouse anti-α-tubulin; clone B-5-1-2, BD Biosciences, Heidelberg, Germany).

Techniques: Expressing, Western Blot, Control

Journal: PLoS ONE

Article Title: Fatty Liver Is Associated with Transcriptional Downregulation of Stearoyl-CoA Desaturase and Impaired Protein Dimerization

doi: 10.1371/journal.pone.0076912

Figure Lengend Snippet: Left panel: Each bar represents the mean ± SE values of Sprague Dawley (SD) rats randomly divided into two experimental groups (standard chow diet CD: n=10; high fat diet HFD: n=15). Right panel: Each bar represents the mean ± SE values of SHR (n=13) and WKY (n=14) divided into two experimental groups (CD:SHR, n= 6 and WKY, n=7; HFD: SHR, n=7 and WKY, n=7). The statistical significance of SHR vs. WKY indicates the comparison between strains independent of diet by two-way ANOVA. In each sample, the Scd1 expression was normalized by the expression of Tbp (TATA box binding protein).

Article Snippet: After blocking with 5% nonfat dry milk in TBST buffer [20 mmol/L of Tris–HCl (pH 7.6), 137 mmol/L of NaCl, and 0.25% of Tween-20], the membranes were probed with rabbit polyclonal anti-Scd1 (Abnova, Taipei, Taiwan), followed by incubation with HRP-conjugated anti-rabbit polyclonal immunoglobulin G secondary antibody (GeneTex, Inc., GTX26795, Irvine, CA, USA).

Techniques: Comparison, Expressing, Binding Assay

Journal: PLoS ONE

Article Title: Fatty Liver Is Associated with Transcriptional Downregulation of Stearoyl-CoA Desaturase and Impaired Protein Dimerization

doi: 10.1371/journal.pone.0076912

Figure Lengend Snippet: Upper left panel: Representative analysis of liver Scd1 protein levels in each experimental model (SHR or WKY) with beta-actin as loading control. Upper right panel: Densitometric quantification of liver SCD1 monomer/dimer ratio protein levels. Values are means ± SE for 13 animals in the SHR group and 14 in the WKY group according to diet, as explained in the legends for and . Lower left and right panels: Densitometric quantification of the liver Scd1 monomeric and dimeric band, respectively, in each experimental group according to diet. Liver scd1 protein expression measured by ELISA. Upper right panel: HFD-induced NAFLD: Each bar represents the mean ± SE values of SD rats randomly divided into two experimental groups (CD: n=10; HFD: n=15). Lower right panel: Each bar represents the mean ± SE values of SHR and WKY divided into two experimental groups (CD:SHR, n= 6 and WKY, n=7; HFD: SHR, n=7 and WKY, n=7). Results are expressed pg/µl proteins.

Article Snippet: After blocking with 5% nonfat dry milk in TBST buffer [20 mmol/L of Tris–HCl (pH 7.6), 137 mmol/L of NaCl, and 0.25% of Tween-20], the membranes were probed with rabbit polyclonal anti-Scd1 (Abnova, Taipei, Taiwan), followed by incubation with HRP-conjugated anti-rabbit polyclonal immunoglobulin G secondary antibody (GeneTex, Inc., GTX26795, Irvine, CA, USA).

Techniques: Control, Expressing, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Fatty Liver Is Associated with Transcriptional Downregulation of Stearoyl-CoA Desaturase and Impaired Protein Dimerization

doi: 10.1371/journal.pone.0076912

Figure Lengend Snippet: SCD (SCD1): stearoyl-CoA desaturase (delta-9-desaturase); CYB5A: cytochrome b5 type A (microsomal); CYP51A1: cytochrome P450, family 51, subfamily A, polypeptide 1, HMGCS1 3-hydroxy-3-methylglutaryl-CoA synthase 1 (soluble); ELOVL5: ELOVL fatty acid elongase 5; SCD5: stearoyl-CoA desaturase 5; FADS6: fatty acid desaturase domain family, member 6; ELOVL2: ELOVL fatty acid elongase 2; FADS1: fatty acid desaturase 1; ELOVL6: ELOVL fatty acid elongase 6; DHCR7: 7-dehydrocholesterol reductase; SQLE: squalene epoxidase; PPARG: peroxisome proliferator-activated receptor gamma; DEGS2: degenerative spermatocyte homolog 2, lipid desaturase; RXRB: retinoid X receptor, beta; RARA: retinoic acid receptor, alpha; TF: transferring; PPARA: peroxisome proliferator-activated receptor alpha; DEGS1: degenerative spermatocyte homolog 1, lipid desaturase; CYB561: cytochrome b-561; PHF20: PHD finger protein 20.

Article Snippet: After blocking with 5% nonfat dry milk in TBST buffer [20 mmol/L of Tris–HCl (pH 7.6), 137 mmol/L of NaCl, and 0.25% of Tween-20], the membranes were probed with rabbit polyclonal anti-Scd1 (Abnova, Taipei, Taiwan), followed by incubation with HRP-conjugated anti-rabbit polyclonal immunoglobulin G secondary antibody (GeneTex, Inc., GTX26795, Irvine, CA, USA).

Techniques: Transferring

Journal: PLoS ONE

Article Title: Fatty Liver Is Associated with Transcriptional Downregulation of Stearoyl-CoA Desaturase and Impaired Protein Dimerization

doi: 10.1371/journal.pone.0076912

Figure Lengend Snippet: Direct SCD1-related nodes are shown in red. A few nodes are labeled for the sake of simplicity. The gene labels are official names.

Article Snippet: After blocking with 5% nonfat dry milk in TBST buffer [20 mmol/L of Tris–HCl (pH 7.6), 137 mmol/L of NaCl, and 0.25% of Tween-20], the membranes were probed with rabbit polyclonal anti-Scd1 (Abnova, Taipei, Taiwan), followed by incubation with HRP-conjugated anti-rabbit polyclonal immunoglobulin G secondary antibody (GeneTex, Inc., GTX26795, Irvine, CA, USA).

Techniques: Labeling

Journal: Cell Systems

Article Title: Context Specificity in Causal Signaling Networks Revealed by Phosphoprotein Profiling

doi: 10.1016/j.cels.2016.11.013

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-SCD1 (clone CD.E10) , Santa Cruz , Cat#sc-58420; RRID: AB_785599.

Techniques: Transduction, Recombinant, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Software, Inhibition, Modification