monoclonal rabbit anti scd1  (Cell Signaling Technology Inc)

Journal: PLOS ONE

Article Title: HIF3A gene disruption causes abnormal alveoli structure and early neonatal death

doi: 10.1371/journal.pone.0300751

Figure Lengend Snippet: (A) Representative images of immunohistochemical staining of SCD1 in neonatal lungs. Scale bar indicates 50 μm. (B) Semiquantitative analysis of SCD1-positive cell counts. Ratios were calculated by dividing SCD1-positive cells from the total cell count. The results are presented as the mean ± SD. Comparisons were analyzed by Kruskal-Wallis test followed by Dunn test. (n = 5). *p < 0.05. (C) Fatty acid composition in lungs of neonatal mice at P0. Total lipids were extracted from lungs and analyzed by gas chromatography. The results are presented as the mean ± SD. All comparisons were analyzed by one-way ANOVA, Tukey’s post hoc test (n = 9 each). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: For immunohistochemistry, deparaffinized sections were incubated overnight at 4°C with rabbit polyclonal anti Ki67 antibody (1:400) (NB500-170; Novus Biologicals, Centennial, CO, USA), rabbit polyclonal anti-PDPN antibody (1:1000) (ab11936; Abcam, Cambridge, UK), rabbit polyclonal anti-Uteroglobin/CC10 antibody (1:2000) (10490-1-AP; Protintech, Rosemont, IL, USA), rabbit polyclonal anti pro SP-C antibody (1:1000)(AB3786; Millipore, Darmstadt, Germany) and rabbit monoclonal anti-SCD1 antibody (1:100) (#2794; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Immunohistochemical staining, Staining, Cell Counting, Gas Chromatography

Journal: Nature cancer

Article Title: Metabolic adaptation of acute lymphoblastic leukemia to the central nervous system microenvironment is dependent on Stearoyl CoA desaturase

doi: 10.1038/s43018-020-00115-2

Figure Lengend Snippet: (A) Quantitative PCR validation of top ranked genes from RNA-Seq in n=5 patient derived xenografts with t(12;21) ETV6-RUNX1 (TEL-AML1) translocation or t(4;11) MLL-AFF1 (MLL-AF4)] translocation. Results are normalized to 36B4 human housekeeping gene and presented as Log2 fold change enrichment comparing CNS to spleen. p (two tailed) = One sample T and Wilcoxon test. (B) Gene expression of SCD1 from RNAseq data deposited in public databases. Left: Samples of BM of patients at diagnosis and relapse and CNS at relapse, unpaired analysis. Right: Patient-derived xenograft samples established by transplantation of patients’ ALL cells onto NSG mice. SCD1 expression in cells isolated from paired CNS and spleen is shown in the graph. FDR – False discovery rate. (C) Intracellular staining of SCD1 in cells from the BM (green) and CNS (grey) of a mouse transplanted with 018z cells. The peaks are relative to the percentage of human CD19+ cells normalized to mode.

Article Snippet: Fifty μg of total protein lysate were loaded on a 12% Polyacrylamide gel and incubated overnight with rabbit anti-human monoclonal SCD1 antibody 1:2000 (SCD1 (C 12 H 5 ) Rabbit mAb #2794, Cell Signaling) and vinculin (Vinculin Antibody #4650, Cell Signaling) at 4°C.

Techniques: Real-time Polymerase Chain Reaction, RNA Sequencing Assay, Derivative Assay, Translocation Assay, Two Tailed Test, Expressing, Transplantation Assay, Isolation, Staining

Journal: Nature cancer

Article Title: Metabolic adaptation of acute lymphoblastic leukemia to the central nervous system microenvironment is dependent on Stearoyl CoA desaturase

doi: 10.1038/s43018-020-00115-2

Figure Lengend Snippet: (A) SCD1 gene expression levels following SCD1 overexpression in 018z cells. (B) Western blot of SCD1 protein after overexpression in 018z cells. (C)I-II Ratio of relative levels of oleoyl-CoA/stearoyl-CoA and of palmitoleoyl-CoA/palmitoyl-CoA in control (CTL) and SCD1-high 018z cells. p=Student’s t-test. (D) Human leukemia burden in 018z xenograft model. Total amount of leukemic cells in CNS, BM and spleen of NSG mice xenografted with human 018z ALL cell line overexpressing SCD1 (SCD1-high) or control (CTL) GFP+ at the time of sacrifice. (E) Human leukemia burden in REH xenograft model. Total amount of leukemic cells in CNS, BM and spleen of NSG mice xenografted with human REH ALL cell lines overexpressing SCD1 (SCD1-high) or control (CTL) GFP+ at the time of sacrifice. p=paired Student’s t-test. (F) Competition assay in vivo: FACS plots of cells injected (INPUT) and cells isolated from the CNS of the mice at sacrifice (OUTPUT). Top: GFP+ control cells transduced with a GFP-carrying lentiviral backbone and mCherry+ control cells transduced with a mCherry carrying lentiviral backbone injected in a ratio of 1:1. Bottom: GFP+ SCD1 overexpressing (SCD1-high) cells and mCherry+ control cells transduced with a mCherry carrying lentiviral backbone injected in a ratio of 1:1. (G) Ratio of total number of SCD1-high 018z cells (GFP+) to control 018z cells (mCherry+) in CNS, BM and spleen of NSG mice transplanted with a mixture of the two cell types in a ratio of 1:1. The backbone vector for SCD1 overexpression was used as control vector. p=paired Student’s t-test.

Article Snippet: Fifty μg of total protein lysate were loaded on a 12% Polyacrylamide gel and incubated overnight with rabbit anti-human monoclonal SCD1 antibody 1:2000 (SCD1 (C 12 H 5 ) Rabbit mAb #2794, Cell Signaling) and vinculin (Vinculin Antibody #4650, Cell Signaling) at 4°C.

Techniques: Expressing, Over Expression, Western Blot, Competitive Binding Assay, In Vivo, Injection, Isolation, Transduction, Plasmid Preparation

Journal: Nature cancer

Article Title: Metabolic adaptation of acute lymphoblastic leukemia to the central nervous system microenvironment is dependent on Stearoyl CoA desaturase

doi: 10.1038/s43018-020-00115-2

Figure Lengend Snippet: (A) Western blot analysis of SCD1 after CRISPR-Cas9 gene ablation in 018z cells. Each lane represents a different gRNA used for the initial screening. gRNA4 decreased SCD1 protein expression while gRNA1-3 did not affect its levels. (B) Gene expression level of SCD1 after CRISPR-Cas9 knockout in 018z cells by gRNA4. (C)I-II Ratio of relative levels of oleoyl-CoA/stearoyl-CoA and of palmitoleoyl-CoA/palmitoyl-CoA in control (CTL) and SCD1-low 018z cells. p=Student’s t-test (D) In vitro proliferation of SCD1-low and control (CTL) 018z cells after 96 hours in in medium supplemented with 10% lipidated or delipidated FBS. The dotted line represents the initial number of cells plated at T0. p=two-way ANOVA. (E) Human leukemia burden in 018z xenograft model. Total amount of leukemic cells in CNS, BM and spleen of NSG mice xenografted with human 018z ALL cell lines SCD1-low or control GFP at the time of sacrifice. p=paired Student’s t-test. (F) Intracellular staining of SCD1 in human cells isolated from the BM of mice transplanted with SCD1-low and control (CTL) 018z cells. The scramble vector for SCD1 downregulation was used as control vector. The peaks are relative to the percentage of human CD19+ cells normalized to mode.

Article Snippet: Fifty μg of total protein lysate were loaded on a 12% Polyacrylamide gel and incubated overnight with rabbit anti-human monoclonal SCD1 antibody 1:2000 (SCD1 (C 12 H 5 ) Rabbit mAb #2794, Cell Signaling) and vinculin (Vinculin Antibody #4650, Cell Signaling) at 4°C.

Techniques: Western Blot, CRISPR, Expressing, Knock-Out, In Vitro, Staining, Isolation, Plasmid Preparation

Journal: Nature cancer

Article Title: Metabolic adaptation of acute lymphoblastic leukemia to the central nervous system microenvironment is dependent on Stearoyl CoA desaturase

doi: 10.1038/s43018-020-00115-2

Figure Lengend Snippet: In vitro proliferation of “SCD1-high”, “SCD1-low” and matching control (CTL) 018z cells after seeding 0.5x106 cells/well for 96 hours in medium supplemented with 10% lipidated (A-B) or delipidated (C-D) FBS treated with 1μM of the SCD1 inhibitor SW203668 or vehicle (DMSO).The dotted line represents the initial number of cells plated at T0. p=two-way ANOVA. (D) GFP+mCherry+FFLuc+ 018z cells were injected intravenously to NSG mice and treated from day 1 to day 10 with the SCD1 inhibitor SW203668 or vehicle (n=5 group). Representative bioluminescence of the tumor load at the time of sacrifice in three different pairs of mice, top: vehicle treated mice; bottom: drug treated mice. Decrease in the tumor load is clear in the spine and the skull area (CNS – marked with white box). Total amount of leukemic cells in CNS (E) and BM (F) of NSG mice xenografted with human GFP+mCherry+FFLuc+ 018z and treated with SW203668 for 10 days at the time of sacrifice. The backbone vector for SCD1 overexpression was used as control vector for SCD1-high and the scramble vector was used as control for SCD1-low. BM – bone marrow. p=Student’s t-test.

Article Snippet: Fifty μg of total protein lysate were loaded on a 12% Polyacrylamide gel and incubated overnight with rabbit anti-human monoclonal SCD1 antibody 1:2000 (SCD1 (C 12 H 5 ) Rabbit mAb #2794, Cell Signaling) and vinculin (Vinculin Antibody #4650, Cell Signaling) at 4°C.

Techniques: In Vitro, Injection, Plasmid Preparation, Over Expression

Journal: Nature cancer

Article Title: Metabolic adaptation of acute lymphoblastic leukemia to the central nervous system microenvironment is dependent on Stearoyl CoA desaturase

doi: 10.1038/s43018-020-00115-2

Figure Lengend Snippet: Human leukemia burden in PDXs. Cells from 4 different PDXs were injected intravenously into NSG mice. Xenografted mice were treated daily from day 7 with the SCD1 inhibitor SW203668 or vehicle (n=5 group) at 20 mg/kg. The total amounts of leukemic cells in CNS and BM of NSG mice at the time of sacrifice are plotted in the graphs (A-D). p=Student’s t-test.

Article Snippet: Fifty μg of total protein lysate were loaded on a 12% Polyacrylamide gel and incubated overnight with rabbit anti-human monoclonal SCD1 antibody 1:2000 (SCD1 (C 12 H 5 ) Rabbit mAb #2794, Cell Signaling) and vinculin (Vinculin Antibody #4650, Cell Signaling) at 4°C.

Techniques: Injection

Journal: PLoS ONE

Article Title: The PPAR pan-agonist tetradecylthioacetic acid promotes redistribution of plasma cholesterol towards large HDL

doi: 10.1371/journal.pone.0229322

Figure Lengend Snippet: a. mRNA expression of Scd1 was analyzed on individual samples (n = 6) with S1 of one individual in the HFD control group used as a calibrator. Data in bar plots are shown as mean ± SEM, in HFD as dark grey bars with individual values in light grey circles and in HFD+TTA as light grey bars with individual values in dark grey circles and student´s unpaired t-test (two tailed) was performed, * = p<0.05 and ** = p<0.01. b. Immunohistochemistry of the middle part of the intestine using an antibody against SCD1. Left panel, HFD control and right panel, HFD+TTA.

Article Snippet: Consecutive sections were incubated overnight with a polyclonal guinea pig anti-mouse Perilipin2 (Progen, GP40) in 1:2000 dilution, and monoclonal rabbit anti-mouse SCD1 (Cell Signaling #2794) in dilution 1:100.

Techniques: Expressing, Control, Two Tailed Test, Immunohistochemistry