recombinant human dll1 expression plasmid  (Sino Biological)


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    Name:
    DLL1 Protein Human Recombinant
    Description:
    A DNA sequence encoding the human DLL1 NP 005609 3 Met1 Gly540 was expressed with the Fc region of human IgG1 at the C terminus
    Catalog Number:
    11635-H02H
    Price:
    None
    Category:
    recombinant protein
    Product Aliases:
    Delta Protein Human, DELTA1 Protein Human, DL1 Protein Human
    Host:
    HEK293 Cells
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    Structured Review

    Sino Biological recombinant human dll1 expression plasmid
    <t>DLL1</t> downregulation decreases migration of MCF-7 and MDA-MB-231 cells and the invasive potential of MDA-MB-231 cells. MCF-7 and MDA-MB-231 cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT) as indicated. (A-B) At 55–70 hours after transfection, cells at 80–90% confluency were scratched, and wound closure was evaluated by microscopy at various time points. Representative images taken at the indicated times post-wounding from three independent experiments are shown. The graph represents mean percentage values (+ SD) of wound closure at each analyzed time point from scratches of these assays. (C) MDA-MB-231 cells were collected 72 hours after transfection and equal cell numbers were added to the upper chamber of 8-μm-pore membranes coated with matrigel and their invasion was measured. Representative fields of crystal-violet-stained cells that invaded to the lower surface of the membranes are shown in each condition. The graphs show mean percentage values (± SD) of invading cells of three independent experiments. *, P
    A DNA sequence encoding the human DLL1 NP 005609 3 Met1 Gly540 was expressed with the Fc region of human IgG1 at the C terminus
    https://www.bioz.com/result/recombinant human dll1 expression plasmid/product/Sino Biological
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant human dll1 expression plasmid - by Bioz Stars, 2021-06
    92/100 stars

    Images

    1) Product Images from "The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells"

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0217002

    DLL1 downregulation decreases migration of MCF-7 and MDA-MB-231 cells and the invasive potential of MDA-MB-231 cells. MCF-7 and MDA-MB-231 cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT) as indicated. (A-B) At 55–70 hours after transfection, cells at 80–90% confluency were scratched, and wound closure was evaluated by microscopy at various time points. Representative images taken at the indicated times post-wounding from three independent experiments are shown. The graph represents mean percentage values (+ SD) of wound closure at each analyzed time point from scratches of these assays. (C) MDA-MB-231 cells were collected 72 hours after transfection and equal cell numbers were added to the upper chamber of 8-μm-pore membranes coated with matrigel and their invasion was measured. Representative fields of crystal-violet-stained cells that invaded to the lower surface of the membranes are shown in each condition. The graphs show mean percentage values (± SD) of invading cells of three independent experiments. *, P
    Figure Legend Snippet: DLL1 downregulation decreases migration of MCF-7 and MDA-MB-231 cells and the invasive potential of MDA-MB-231 cells. MCF-7 and MDA-MB-231 cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT) as indicated. (A-B) At 55–70 hours after transfection, cells at 80–90% confluency were scratched, and wound closure was evaluated by microscopy at various time points. Representative images taken at the indicated times post-wounding from three independent experiments are shown. The graph represents mean percentage values (+ SD) of wound closure at each analyzed time point from scratches of these assays. (C) MDA-MB-231 cells were collected 72 hours after transfection and equal cell numbers were added to the upper chamber of 8-μm-pore membranes coated with matrigel and their invasion was measured. Representative fields of crystal-violet-stained cells that invaded to the lower surface of the membranes are shown in each condition. The graphs show mean percentage values (± SD) of invading cells of three independent experiments. *, P

    Techniques Used: Migration, Multiple Displacement Amplification, Transfection, Negative Control, Microscopy, Staining

    DLL1 downregulation in MCF-7 cells decreases proliferation and induces cell cycle arrest in the G1 phase. MCF-7, BT474, and MDA-MB-231 cells were not transfected (NT) or transfected with DLL1-siRNA (DLL1-si1/2), or negative control (Ctr) siRNA as indicated. (A-B) Cellular growth was determined by microscopy using the trypan blue exclusion (A) and the MTT (B) methods at 72 hours after transfection in MCF-7 and 72 up to120 hours in BT474 and MDA-MB-231 cells. Values were calculated as relative percentage of cells transfected with Ctr-siRNA, in each assay for each cell type. The graphs represent data (mean ± SD) from at least three independent assays. (C-E) At 86 hours post-transfection cells were collected, fixed, stained with propidium iodide, and DNA content was analyzed by flow cytometry. Representative histograms showing the effect of DLL1 downregulation in cell cycle progression from these assays are shown. Graph shows mean percentage (+ SD) of cells in each phase of the cell cycle of four independent assays. *, P
    Figure Legend Snippet: DLL1 downregulation in MCF-7 cells decreases proliferation and induces cell cycle arrest in the G1 phase. MCF-7, BT474, and MDA-MB-231 cells were not transfected (NT) or transfected with DLL1-siRNA (DLL1-si1/2), or negative control (Ctr) siRNA as indicated. (A-B) Cellular growth was determined by microscopy using the trypan blue exclusion (A) and the MTT (B) methods at 72 hours after transfection in MCF-7 and 72 up to120 hours in BT474 and MDA-MB-231 cells. Values were calculated as relative percentage of cells transfected with Ctr-siRNA, in each assay for each cell type. The graphs represent data (mean ± SD) from at least three independent assays. (C-E) At 86 hours post-transfection cells were collected, fixed, stained with propidium iodide, and DNA content was analyzed by flow cytometry. Representative histograms showing the effect of DLL1 downregulation in cell cycle progression from these assays are shown. Graph shows mean percentage (+ SD) of cells in each phase of the cell cycle of four independent assays. *, P

    Techniques Used: Multiple Displacement Amplification, Transfection, Negative Control, Microscopy, MTT Assay, Staining, Flow Cytometry

    DLL1 downregulation impairs Notch pathway activation in BC cells and decreases MCF-7 and BT474 colony formation abilities. (A) Expression of DLL1 mRNA and protein levels in luminal A MCF-7, luminal B BT474, and triple-negative MDA-MB-231 cells were quantitated by qRT-PCR and immunoblotting. The DLL1 mRNA values were normalized against the HPRT1 mRNA levels in the same samples. α-tubulin was used as the loading control. (B-F) Cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT). (B) Total RNA was extracted from the indicated cells 24–38 hours post-transfection and DLL1 mRNA levels were quantitated by qRT-PCR. (C) Total soluble protein extracts were prepared from cells 72 hours following transfection and the levels of DLL1 protein were assessed by immunoblotting. The numbers under the bands indicate DLL1 fold changes relative to Ctr-siRNA transfected cells after normalization against α-tubulin. (D) qRT-PCR analysis of HEY1 mRNA in the indicated cells at 48–72 hours after transfection. The values in (B) and (D) were normalized against the HPRT1 mRNA levels in the same sample and calculated as mean fold change (+ SD) relative to the respective control cells transfected with Ctr-siRNA. Graphs in (A, B and D) represents mean (+ SD) of at least three independent assays. (E-F) Colony formation of MCF-7, BT474 and MDA-MB-231 cells at day 9 post transfection. Representative fields of crystal-violet-stained colonies for each cell type of four independent assays are shown. The graphs show mean percentage of the number of colonies (+ SD) relative to the respective control cells transfected with Ctr-siRNA from these assays. *, P
    Figure Legend Snippet: DLL1 downregulation impairs Notch pathway activation in BC cells and decreases MCF-7 and BT474 colony formation abilities. (A) Expression of DLL1 mRNA and protein levels in luminal A MCF-7, luminal B BT474, and triple-negative MDA-MB-231 cells were quantitated by qRT-PCR and immunoblotting. The DLL1 mRNA values were normalized against the HPRT1 mRNA levels in the same samples. α-tubulin was used as the loading control. (B-F) Cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT). (B) Total RNA was extracted from the indicated cells 24–38 hours post-transfection and DLL1 mRNA levels were quantitated by qRT-PCR. (C) Total soluble protein extracts were prepared from cells 72 hours following transfection and the levels of DLL1 protein were assessed by immunoblotting. The numbers under the bands indicate DLL1 fold changes relative to Ctr-siRNA transfected cells after normalization against α-tubulin. (D) qRT-PCR analysis of HEY1 mRNA in the indicated cells at 48–72 hours after transfection. The values in (B) and (D) were normalized against the HPRT1 mRNA levels in the same sample and calculated as mean fold change (+ SD) relative to the respective control cells transfected with Ctr-siRNA. Graphs in (A, B and D) represents mean (+ SD) of at least three independent assays. (E-F) Colony formation of MCF-7, BT474 and MDA-MB-231 cells at day 9 post transfection. Representative fields of crystal-violet-stained colonies for each cell type of four independent assays are shown. The graphs show mean percentage of the number of colonies (+ SD) relative to the respective control cells transfected with Ctr-siRNA from these assays. *, P

    Techniques Used: Activation Assay, Expressing, Multiple Displacement Amplification, Quantitative RT-PCR, Transfection, Negative Control, Staining

    Human recombinant DLL1 activates the Notch pathway and promotes MCF-7 cell proliferation and migration. (A) MCF-7 cells were plated in tissue culture plates not coated (control) or pre-coated with DLL1-Fc or Fc proteins in the absence (vehicle) or the presence of the Notch pathway inhibitor DAPT. Total RNA was extracted 17 hours thereafter and the expression levels of the indicated Notch-target genes were quantified by qRT-PCR, and calculated as fold change relative to control non-treated cells. Graph represents data (mean + SD) from three independent experiments. *, P
    Figure Legend Snippet: Human recombinant DLL1 activates the Notch pathway and promotes MCF-7 cell proliferation and migration. (A) MCF-7 cells were plated in tissue culture plates not coated (control) or pre-coated with DLL1-Fc or Fc proteins in the absence (vehicle) or the presence of the Notch pathway inhibitor DAPT. Total RNA was extracted 17 hours thereafter and the expression levels of the indicated Notch-target genes were quantified by qRT-PCR, and calculated as fold change relative to control non-treated cells. Graph represents data (mean + SD) from three independent experiments. *, P

    Techniques Used: Recombinant, Migration, Expressing, Quantitative RT-PCR

    DLL1 downregulation promotes apoptosis of MCF-7 cells. (A-C) MCF-7, BT474 and MDA-MB-231 cells were not-transfected (NT) or transfected with DLL1-siRNAs (DLL1-si1/2), or negative control (Ctr) siRNA. At 92 hours (MCF-7) and 120 hours (BT474 and MDA-MB-231) after transfection cells were collected, stained with annexin-V and apoptosis analyzed by flow cytometry. Representative dot plots showing the percentage of apoptosis at each condition are represented. Graph shows percentage of apoptosis (mean + SD) of at least three independent assays, each one performed in triplicate. *, P
    Figure Legend Snippet: DLL1 downregulation promotes apoptosis of MCF-7 cells. (A-C) MCF-7, BT474 and MDA-MB-231 cells were not-transfected (NT) or transfected with DLL1-siRNAs (DLL1-si1/2), or negative control (Ctr) siRNA. At 92 hours (MCF-7) and 120 hours (BT474 and MDA-MB-231) after transfection cells were collected, stained with annexin-V and apoptosis analyzed by flow cytometry. Representative dot plots showing the percentage of apoptosis at each condition are represented. Graph shows percentage of apoptosis (mean + SD) of at least three independent assays, each one performed in triplicate. *, P

    Techniques Used: Multiple Displacement Amplification, Transfection, Negative Control, Staining, Flow Cytometry

    DLL1 downregulation in MCF-7 cells upregulate genes involved in cell cycle arrest and represses genes that promote cell cycle progression, proliferation and survival. Cells were transfected with DLL1-siRNA1 or Ctr-siRNA. (A) At 55–70 hours following transfection total RNA was extracted and the expression of genes that control cell cycle arrest (1), cell cycle transition (2), proliferation (3), and survival (4) were quantified by qRT-PCR. The values were normalized against the HPRT1 mRNA levels in the same sample and calculated as fold change relative to control cells transfected with Ctr-siRNA. Results are represented as mean values (+ SD) from four independent assays. (B) The expression levels of the indicated proteins were determined by immunoblotting 72 hours after transfection in total soluble protein extracts. α-tubulin was used as an internal control. The graph shows relative expression of these proteins in cells transfected with DLL1-siRNA1 as compared to Ctr-siRNA transfected cells of at least three independent experiments. *, P
    Figure Legend Snippet: DLL1 downregulation in MCF-7 cells upregulate genes involved in cell cycle arrest and represses genes that promote cell cycle progression, proliferation and survival. Cells were transfected with DLL1-siRNA1 or Ctr-siRNA. (A) At 55–70 hours following transfection total RNA was extracted and the expression of genes that control cell cycle arrest (1), cell cycle transition (2), proliferation (3), and survival (4) were quantified by qRT-PCR. The values were normalized against the HPRT1 mRNA levels in the same sample and calculated as fold change relative to control cells transfected with Ctr-siRNA. Results are represented as mean values (+ SD) from four independent assays. (B) The expression levels of the indicated proteins were determined by immunoblotting 72 hours after transfection in total soluble protein extracts. α-tubulin was used as an internal control. The graph shows relative expression of these proteins in cells transfected with DLL1-siRNA1 as compared to Ctr-siRNA transfected cells of at least three independent experiments. *, P

    Techniques Used: Transfection, Expressing, Quantitative RT-PCR

    Related Articles

    Recombinant:

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    Article Snippet: Cells, Proteins, Plasmids, and Other ReagentsWe purchased 293 FreeStyle (293FS) cells and protein A agarose from Thermo Fisher Scientific. .. Recombinant human (hHER2), cynomolgus (cHER2), mouse (mHER2), and rat HER2 (rHER2) and other ErbB family member proteins were purchased from Sino Biological (Beijing, China). .. The pWC1 vector for phage display and bacterial expression and the pDin1 vector used for mammalian expression were designed and generated in our laboratory.

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    Article Snippet: .. We evaluated the use of gold nanoparticles (AuNPs) platform in a homogenous assay for a potency measurement of a therapeutic monoclonal antibody (mAb). .. We evaluated the use of gold nanoparticles (AuNPs) platform in a homogenous assay for a potency measurement of a therapeutic monoclonal antibody (mAb).

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells
    Article Snippet: The Notch pathway signaling inhibitor DAPT (Sigma, D5942) was used at 5 μM. .. Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ . .. The amplified cDNA was cloned into the mammalian pFUSE-IgG1-Fc vector (InvivoGen, pfuse-hg1fc1, Toulouse, France) in frame with Fc tag at the C-terminal region, using standard techniques.

    Expressing:

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells
    Article Snippet: The Notch pathway signaling inhibitor DAPT (Sigma, D5942) was used at 5 μM. .. Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ . .. The amplified cDNA was cloned into the mammalian pFUSE-IgG1-Fc vector (InvivoGen, pfuse-hg1fc1, Toulouse, France) in frame with Fc tag at the C-terminal region, using standard techniques.

    Plasmid Preparation:

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells
    Article Snippet: The Notch pathway signaling inhibitor DAPT (Sigma, D5942) was used at 5 μM. .. Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ . .. The amplified cDNA was cloned into the mammalian pFUSE-IgG1-Fc vector (InvivoGen, pfuse-hg1fc1, Toulouse, France) in frame with Fc tag at the C-terminal region, using standard techniques.

    Purification:

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells
    Article Snippet: The Notch pathway signaling inhibitor DAPT (Sigma, D5942) was used at 5 μM. .. Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ . .. The amplified cDNA was cloned into the mammalian pFUSE-IgG1-Fc vector (InvivoGen, pfuse-hg1fc1, Toulouse, France) in frame with Fc tag at the C-terminal region, using standard techniques.

    Amplification:

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells
    Article Snippet: The Notch pathway signaling inhibitor DAPT (Sigma, D5942) was used at 5 μM. .. Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ . .. The amplified cDNA was cloned into the mammalian pFUSE-IgG1-Fc vector (InvivoGen, pfuse-hg1fc1, Toulouse, France) in frame with Fc tag at the C-terminal region, using standard techniques.

    Polymerase Chain Reaction:

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells
    Article Snippet: The Notch pathway signaling inhibitor DAPT (Sigma, D5942) was used at 5 μM. .. Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ . .. The amplified cDNA was cloned into the mammalian pFUSE-IgG1-Fc vector (InvivoGen, pfuse-hg1fc1, Toulouse, France) in frame with Fc tag at the C-terminal region, using standard techniques.

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  • 92
    Sino Biological human dll1 signal peptide
    <t>DLL1</t> downregulation decreases migration of MCF-7 and MDA-MB-231 cells and the invasive potential of MDA-MB-231 cells. MCF-7 and MDA-MB-231 cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT) as indicated. (A-B) At 55–70 hours after transfection, cells at 80–90% confluency were scratched, and wound closure was evaluated by microscopy at various time points. Representative images taken at the indicated times post-wounding from three independent experiments are shown. The graph represents mean percentage values (+ SD) of wound closure at each analyzed time point from scratches of these assays. (C) MDA-MB-231 cells were collected 72 hours after transfection and equal cell numbers were added to the upper chamber of 8-μm-pore membranes coated with matrigel and their invasion was measured. Representative fields of crystal-violet-stained cells that invaded to the lower surface of the membranes are shown in each condition. The graphs show mean percentage values (± SD) of invading cells of three independent experiments. *, P
    Human Dll1 Signal Peptide, supplied by Sino Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human dll1 signal peptide/product/Sino Biological
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human dll1 signal peptide - by Bioz Stars, 2021-06
    92/100 stars
      Buy from Supplier

    92
    Sino Biological recombinant human dll1 expression plasmid
    <t>DLL1</t> downregulation decreases migration of MCF-7 and MDA-MB-231 cells and the invasive potential of MDA-MB-231 cells. MCF-7 and MDA-MB-231 cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT) as indicated. (A-B) At 55–70 hours after transfection, cells at 80–90% confluency were scratched, and wound closure was evaluated by microscopy at various time points. Representative images taken at the indicated times post-wounding from three independent experiments are shown. The graph represents mean percentage values (+ SD) of wound closure at each analyzed time point from scratches of these assays. (C) MDA-MB-231 cells were collected 72 hours after transfection and equal cell numbers were added to the upper chamber of 8-μm-pore membranes coated with matrigel and their invasion was measured. Representative fields of crystal-violet-stained cells that invaded to the lower surface of the membranes are shown in each condition. The graphs show mean percentage values (± SD) of invading cells of three independent experiments. *, P
    Recombinant Human Dll1 Expression Plasmid, supplied by Sino Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant human dll1 expression plasmid/product/Sino Biological
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant human dll1 expression plasmid - by Bioz Stars, 2021-06
    92/100 stars
      Buy from Supplier

    Image Search Results


    DLL1 downregulation decreases migration of MCF-7 and MDA-MB-231 cells and the invasive potential of MDA-MB-231 cells. MCF-7 and MDA-MB-231 cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT) as indicated. (A-B) At 55–70 hours after transfection, cells at 80–90% confluency were scratched, and wound closure was evaluated by microscopy at various time points. Representative images taken at the indicated times post-wounding from three independent experiments are shown. The graph represents mean percentage values (+ SD) of wound closure at each analyzed time point from scratches of these assays. (C) MDA-MB-231 cells were collected 72 hours after transfection and equal cell numbers were added to the upper chamber of 8-μm-pore membranes coated with matrigel and their invasion was measured. Representative fields of crystal-violet-stained cells that invaded to the lower surface of the membranes are shown in each condition. The graphs show mean percentage values (± SD) of invading cells of three independent experiments. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation decreases migration of MCF-7 and MDA-MB-231 cells and the invasive potential of MDA-MB-231 cells. MCF-7 and MDA-MB-231 cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT) as indicated. (A-B) At 55–70 hours after transfection, cells at 80–90% confluency were scratched, and wound closure was evaluated by microscopy at various time points. Representative images taken at the indicated times post-wounding from three independent experiments are shown. The graph represents mean percentage values (+ SD) of wound closure at each analyzed time point from scratches of these assays. (C) MDA-MB-231 cells were collected 72 hours after transfection and equal cell numbers were added to the upper chamber of 8-μm-pore membranes coated with matrigel and their invasion was measured. Representative fields of crystal-violet-stained cells that invaded to the lower surface of the membranes are shown in each condition. The graphs show mean percentage values (± SD) of invading cells of three independent experiments. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Migration, Multiple Displacement Amplification, Transfection, Negative Control, Microscopy, Staining

    DLL1 downregulation in MCF-7 cells decreases proliferation and induces cell cycle arrest in the G1 phase. MCF-7, BT474, and MDA-MB-231 cells were not transfected (NT) or transfected with DLL1-siRNA (DLL1-si1/2), or negative control (Ctr) siRNA as indicated. (A-B) Cellular growth was determined by microscopy using the trypan blue exclusion (A) and the MTT (B) methods at 72 hours after transfection in MCF-7 and 72 up to120 hours in BT474 and MDA-MB-231 cells. Values were calculated as relative percentage of cells transfected with Ctr-siRNA, in each assay for each cell type. The graphs represent data (mean ± SD) from at least three independent assays. (C-E) At 86 hours post-transfection cells were collected, fixed, stained with propidium iodide, and DNA content was analyzed by flow cytometry. Representative histograms showing the effect of DLL1 downregulation in cell cycle progression from these assays are shown. Graph shows mean percentage (+ SD) of cells in each phase of the cell cycle of four independent assays. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation in MCF-7 cells decreases proliferation and induces cell cycle arrest in the G1 phase. MCF-7, BT474, and MDA-MB-231 cells were not transfected (NT) or transfected with DLL1-siRNA (DLL1-si1/2), or negative control (Ctr) siRNA as indicated. (A-B) Cellular growth was determined by microscopy using the trypan blue exclusion (A) and the MTT (B) methods at 72 hours after transfection in MCF-7 and 72 up to120 hours in BT474 and MDA-MB-231 cells. Values were calculated as relative percentage of cells transfected with Ctr-siRNA, in each assay for each cell type. The graphs represent data (mean ± SD) from at least three independent assays. (C-E) At 86 hours post-transfection cells were collected, fixed, stained with propidium iodide, and DNA content was analyzed by flow cytometry. Representative histograms showing the effect of DLL1 downregulation in cell cycle progression from these assays are shown. Graph shows mean percentage (+ SD) of cells in each phase of the cell cycle of four independent assays. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Multiple Displacement Amplification, Transfection, Negative Control, Microscopy, MTT Assay, Staining, Flow Cytometry

    DLL1 downregulation impairs Notch pathway activation in BC cells and decreases MCF-7 and BT474 colony formation abilities. (A) Expression of DLL1 mRNA and protein levels in luminal A MCF-7, luminal B BT474, and triple-negative MDA-MB-231 cells were quantitated by qRT-PCR and immunoblotting. The DLL1 mRNA values were normalized against the HPRT1 mRNA levels in the same samples. α-tubulin was used as the loading control. (B-F) Cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT). (B) Total RNA was extracted from the indicated cells 24–38 hours post-transfection and DLL1 mRNA levels were quantitated by qRT-PCR. (C) Total soluble protein extracts were prepared from cells 72 hours following transfection and the levels of DLL1 protein were assessed by immunoblotting. The numbers under the bands indicate DLL1 fold changes relative to Ctr-siRNA transfected cells after normalization against α-tubulin. (D) qRT-PCR analysis of HEY1 mRNA in the indicated cells at 48–72 hours after transfection. The values in (B) and (D) were normalized against the HPRT1 mRNA levels in the same sample and calculated as mean fold change (+ SD) relative to the respective control cells transfected with Ctr-siRNA. Graphs in (A, B and D) represents mean (+ SD) of at least three independent assays. (E-F) Colony formation of MCF-7, BT474 and MDA-MB-231 cells at day 9 post transfection. Representative fields of crystal-violet-stained colonies for each cell type of four independent assays are shown. The graphs show mean percentage of the number of colonies (+ SD) relative to the respective control cells transfected with Ctr-siRNA from these assays. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation impairs Notch pathway activation in BC cells and decreases MCF-7 and BT474 colony formation abilities. (A) Expression of DLL1 mRNA and protein levels in luminal A MCF-7, luminal B BT474, and triple-negative MDA-MB-231 cells were quantitated by qRT-PCR and immunoblotting. The DLL1 mRNA values were normalized against the HPRT1 mRNA levels in the same samples. α-tubulin was used as the loading control. (B-F) Cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT). (B) Total RNA was extracted from the indicated cells 24–38 hours post-transfection and DLL1 mRNA levels were quantitated by qRT-PCR. (C) Total soluble protein extracts were prepared from cells 72 hours following transfection and the levels of DLL1 protein were assessed by immunoblotting. The numbers under the bands indicate DLL1 fold changes relative to Ctr-siRNA transfected cells after normalization against α-tubulin. (D) qRT-PCR analysis of HEY1 mRNA in the indicated cells at 48–72 hours after transfection. The values in (B) and (D) were normalized against the HPRT1 mRNA levels in the same sample and calculated as mean fold change (+ SD) relative to the respective control cells transfected with Ctr-siRNA. Graphs in (A, B and D) represents mean (+ SD) of at least three independent assays. (E-F) Colony formation of MCF-7, BT474 and MDA-MB-231 cells at day 9 post transfection. Representative fields of crystal-violet-stained colonies for each cell type of four independent assays are shown. The graphs show mean percentage of the number of colonies (+ SD) relative to the respective control cells transfected with Ctr-siRNA from these assays. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Activation Assay, Expressing, Multiple Displacement Amplification, Quantitative RT-PCR, Transfection, Negative Control, Staining

    Human recombinant DLL1 activates the Notch pathway and promotes MCF-7 cell proliferation and migration. (A) MCF-7 cells were plated in tissue culture plates not coated (control) or pre-coated with DLL1-Fc or Fc proteins in the absence (vehicle) or the presence of the Notch pathway inhibitor DAPT. Total RNA was extracted 17 hours thereafter and the expression levels of the indicated Notch-target genes were quantified by qRT-PCR, and calculated as fold change relative to control non-treated cells. Graph represents data (mean + SD) from three independent experiments. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: Human recombinant DLL1 activates the Notch pathway and promotes MCF-7 cell proliferation and migration. (A) MCF-7 cells were plated in tissue culture plates not coated (control) or pre-coated with DLL1-Fc or Fc proteins in the absence (vehicle) or the presence of the Notch pathway inhibitor DAPT. Total RNA was extracted 17 hours thereafter and the expression levels of the indicated Notch-target genes were quantified by qRT-PCR, and calculated as fold change relative to control non-treated cells. Graph represents data (mean + SD) from three independent experiments. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Recombinant, Migration, Expressing, Quantitative RT-PCR

    DLL1 downregulation promotes apoptosis of MCF-7 cells. (A-C) MCF-7, BT474 and MDA-MB-231 cells were not-transfected (NT) or transfected with DLL1-siRNAs (DLL1-si1/2), or negative control (Ctr) siRNA. At 92 hours (MCF-7) and 120 hours (BT474 and MDA-MB-231) after transfection cells were collected, stained with annexin-V and apoptosis analyzed by flow cytometry. Representative dot plots showing the percentage of apoptosis at each condition are represented. Graph shows percentage of apoptosis (mean + SD) of at least three independent assays, each one performed in triplicate. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation promotes apoptosis of MCF-7 cells. (A-C) MCF-7, BT474 and MDA-MB-231 cells were not-transfected (NT) or transfected with DLL1-siRNAs (DLL1-si1/2), or negative control (Ctr) siRNA. At 92 hours (MCF-7) and 120 hours (BT474 and MDA-MB-231) after transfection cells were collected, stained with annexin-V and apoptosis analyzed by flow cytometry. Representative dot plots showing the percentage of apoptosis at each condition are represented. Graph shows percentage of apoptosis (mean + SD) of at least three independent assays, each one performed in triplicate. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Multiple Displacement Amplification, Transfection, Negative Control, Staining, Flow Cytometry

    DLL1 downregulation in MCF-7 cells upregulate genes involved in cell cycle arrest and represses genes that promote cell cycle progression, proliferation and survival. Cells were transfected with DLL1-siRNA1 or Ctr-siRNA. (A) At 55–70 hours following transfection total RNA was extracted and the expression of genes that control cell cycle arrest (1), cell cycle transition (2), proliferation (3), and survival (4) were quantified by qRT-PCR. The values were normalized against the HPRT1 mRNA levels in the same sample and calculated as fold change relative to control cells transfected with Ctr-siRNA. Results are represented as mean values (+ SD) from four independent assays. (B) The expression levels of the indicated proteins were determined by immunoblotting 72 hours after transfection in total soluble protein extracts. α-tubulin was used as an internal control. The graph shows relative expression of these proteins in cells transfected with DLL1-siRNA1 as compared to Ctr-siRNA transfected cells of at least three independent experiments. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation in MCF-7 cells upregulate genes involved in cell cycle arrest and represses genes that promote cell cycle progression, proliferation and survival. Cells were transfected with DLL1-siRNA1 or Ctr-siRNA. (A) At 55–70 hours following transfection total RNA was extracted and the expression of genes that control cell cycle arrest (1), cell cycle transition (2), proliferation (3), and survival (4) were quantified by qRT-PCR. The values were normalized against the HPRT1 mRNA levels in the same sample and calculated as fold change relative to control cells transfected with Ctr-siRNA. Results are represented as mean values (+ SD) from four independent assays. (B) The expression levels of the indicated proteins were determined by immunoblotting 72 hours after transfection in total soluble protein extracts. α-tubulin was used as an internal control. The graph shows relative expression of these proteins in cells transfected with DLL1-siRNA1 as compared to Ctr-siRNA transfected cells of at least three independent experiments. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Transfection, Expressing, Quantitative RT-PCR

    DLL1 downregulation decreases migration of MCF-7 and MDA-MB-231 cells and the invasive potential of MDA-MB-231 cells. MCF-7 and MDA-MB-231 cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT) as indicated. (A-B) At 55–70 hours after transfection, cells at 80–90% confluency were scratched, and wound closure was evaluated by microscopy at various time points. Representative images taken at the indicated times post-wounding from three independent experiments are shown. The graph represents mean percentage values (+ SD) of wound closure at each analyzed time point from scratches of these assays. (C) MDA-MB-231 cells were collected 72 hours after transfection and equal cell numbers were added to the upper chamber of 8-μm-pore membranes coated with matrigel and their invasion was measured. Representative fields of crystal-violet-stained cells that invaded to the lower surface of the membranes are shown in each condition. The graphs show mean percentage values (± SD) of invading cells of three independent experiments. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation decreases migration of MCF-7 and MDA-MB-231 cells and the invasive potential of MDA-MB-231 cells. MCF-7 and MDA-MB-231 cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT) as indicated. (A-B) At 55–70 hours after transfection, cells at 80–90% confluency were scratched, and wound closure was evaluated by microscopy at various time points. Representative images taken at the indicated times post-wounding from three independent experiments are shown. The graph represents mean percentage values (+ SD) of wound closure at each analyzed time point from scratches of these assays. (C) MDA-MB-231 cells were collected 72 hours after transfection and equal cell numbers were added to the upper chamber of 8-μm-pore membranes coated with matrigel and their invasion was measured. Representative fields of crystal-violet-stained cells that invaded to the lower surface of the membranes are shown in each condition. The graphs show mean percentage values (± SD) of invading cells of three independent experiments. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Migration, Multiple Displacement Amplification, Transfection, Negative Control, Microscopy, Staining

    DLL1 downregulation in MCF-7 cells decreases proliferation and induces cell cycle arrest in the G1 phase. MCF-7, BT474, and MDA-MB-231 cells were not transfected (NT) or transfected with DLL1-siRNA (DLL1-si1/2), or negative control (Ctr) siRNA as indicated. (A-B) Cellular growth was determined by microscopy using the trypan blue exclusion (A) and the MTT (B) methods at 72 hours after transfection in MCF-7 and 72 up to120 hours in BT474 and MDA-MB-231 cells. Values were calculated as relative percentage of cells transfected with Ctr-siRNA, in each assay for each cell type. The graphs represent data (mean ± SD) from at least three independent assays. (C-E) At 86 hours post-transfection cells were collected, fixed, stained with propidium iodide, and DNA content was analyzed by flow cytometry. Representative histograms showing the effect of DLL1 downregulation in cell cycle progression from these assays are shown. Graph shows mean percentage (+ SD) of cells in each phase of the cell cycle of four independent assays. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation in MCF-7 cells decreases proliferation and induces cell cycle arrest in the G1 phase. MCF-7, BT474, and MDA-MB-231 cells were not transfected (NT) or transfected with DLL1-siRNA (DLL1-si1/2), or negative control (Ctr) siRNA as indicated. (A-B) Cellular growth was determined by microscopy using the trypan blue exclusion (A) and the MTT (B) methods at 72 hours after transfection in MCF-7 and 72 up to120 hours in BT474 and MDA-MB-231 cells. Values were calculated as relative percentage of cells transfected with Ctr-siRNA, in each assay for each cell type. The graphs represent data (mean ± SD) from at least three independent assays. (C-E) At 86 hours post-transfection cells were collected, fixed, stained with propidium iodide, and DNA content was analyzed by flow cytometry. Representative histograms showing the effect of DLL1 downregulation in cell cycle progression from these assays are shown. Graph shows mean percentage (+ SD) of cells in each phase of the cell cycle of four independent assays. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Multiple Displacement Amplification, Transfection, Negative Control, Microscopy, MTT Assay, Staining, Flow Cytometry

    DLL1 downregulation impairs Notch pathway activation in BC cells and decreases MCF-7 and BT474 colony formation abilities. (A) Expression of DLL1 mRNA and protein levels in luminal A MCF-7, luminal B BT474, and triple-negative MDA-MB-231 cells were quantitated by qRT-PCR and immunoblotting. The DLL1 mRNA values were normalized against the HPRT1 mRNA levels in the same samples. α-tubulin was used as the loading control. (B-F) Cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT). (B) Total RNA was extracted from the indicated cells 24–38 hours post-transfection and DLL1 mRNA levels were quantitated by qRT-PCR. (C) Total soluble protein extracts were prepared from cells 72 hours following transfection and the levels of DLL1 protein were assessed by immunoblotting. The numbers under the bands indicate DLL1 fold changes relative to Ctr-siRNA transfected cells after normalization against α-tubulin. (D) qRT-PCR analysis of HEY1 mRNA in the indicated cells at 48–72 hours after transfection. The values in (B) and (D) were normalized against the HPRT1 mRNA levels in the same sample and calculated as mean fold change (+ SD) relative to the respective control cells transfected with Ctr-siRNA. Graphs in (A, B and D) represents mean (+ SD) of at least three independent assays. (E-F) Colony formation of MCF-7, BT474 and MDA-MB-231 cells at day 9 post transfection. Representative fields of crystal-violet-stained colonies for each cell type of four independent assays are shown. The graphs show mean percentage of the number of colonies (+ SD) relative to the respective control cells transfected with Ctr-siRNA from these assays. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation impairs Notch pathway activation in BC cells and decreases MCF-7 and BT474 colony formation abilities. (A) Expression of DLL1 mRNA and protein levels in luminal A MCF-7, luminal B BT474, and triple-negative MDA-MB-231 cells were quantitated by qRT-PCR and immunoblotting. The DLL1 mRNA values were normalized against the HPRT1 mRNA levels in the same samples. α-tubulin was used as the loading control. (B-F) Cells were transiently transfected with DLL1 siRNAs (DLL1-si1/2), negative control siRNA (Ctr-si) or not-transfected (NT). (B) Total RNA was extracted from the indicated cells 24–38 hours post-transfection and DLL1 mRNA levels were quantitated by qRT-PCR. (C) Total soluble protein extracts were prepared from cells 72 hours following transfection and the levels of DLL1 protein were assessed by immunoblotting. The numbers under the bands indicate DLL1 fold changes relative to Ctr-siRNA transfected cells after normalization against α-tubulin. (D) qRT-PCR analysis of HEY1 mRNA in the indicated cells at 48–72 hours after transfection. The values in (B) and (D) were normalized against the HPRT1 mRNA levels in the same sample and calculated as mean fold change (+ SD) relative to the respective control cells transfected with Ctr-siRNA. Graphs in (A, B and D) represents mean (+ SD) of at least three independent assays. (E-F) Colony formation of MCF-7, BT474 and MDA-MB-231 cells at day 9 post transfection. Representative fields of crystal-violet-stained colonies for each cell type of four independent assays are shown. The graphs show mean percentage of the number of colonies (+ SD) relative to the respective control cells transfected with Ctr-siRNA from these assays. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Activation Assay, Expressing, Multiple Displacement Amplification, Quantitative RT-PCR, Transfection, Negative Control, Staining

    Human recombinant DLL1 activates the Notch pathway and promotes MCF-7 cell proliferation and migration. (A) MCF-7 cells were plated in tissue culture plates not coated (control) or pre-coated with DLL1-Fc or Fc proteins in the absence (vehicle) or the presence of the Notch pathway inhibitor DAPT. Total RNA was extracted 17 hours thereafter and the expression levels of the indicated Notch-target genes were quantified by qRT-PCR, and calculated as fold change relative to control non-treated cells. Graph represents data (mean + SD) from three independent experiments. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: Human recombinant DLL1 activates the Notch pathway and promotes MCF-7 cell proliferation and migration. (A) MCF-7 cells were plated in tissue culture plates not coated (control) or pre-coated with DLL1-Fc or Fc proteins in the absence (vehicle) or the presence of the Notch pathway inhibitor DAPT. Total RNA was extracted 17 hours thereafter and the expression levels of the indicated Notch-target genes were quantified by qRT-PCR, and calculated as fold change relative to control non-treated cells. Graph represents data (mean + SD) from three independent experiments. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Recombinant, Migration, Expressing, Quantitative RT-PCR

    DLL1 downregulation promotes apoptosis of MCF-7 cells. (A-C) MCF-7, BT474 and MDA-MB-231 cells were not-transfected (NT) or transfected with DLL1-siRNAs (DLL1-si1/2), or negative control (Ctr) siRNA. At 92 hours (MCF-7) and 120 hours (BT474 and MDA-MB-231) after transfection cells were collected, stained with annexin-V and apoptosis analyzed by flow cytometry. Representative dot plots showing the percentage of apoptosis at each condition are represented. Graph shows percentage of apoptosis (mean + SD) of at least three independent assays, each one performed in triplicate. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation promotes apoptosis of MCF-7 cells. (A-C) MCF-7, BT474 and MDA-MB-231 cells were not-transfected (NT) or transfected with DLL1-siRNAs (DLL1-si1/2), or negative control (Ctr) siRNA. At 92 hours (MCF-7) and 120 hours (BT474 and MDA-MB-231) after transfection cells were collected, stained with annexin-V and apoptosis analyzed by flow cytometry. Representative dot plots showing the percentage of apoptosis at each condition are represented. Graph shows percentage of apoptosis (mean + SD) of at least three independent assays, each one performed in triplicate. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Multiple Displacement Amplification, Transfection, Negative Control, Staining, Flow Cytometry

    DLL1 downregulation in MCF-7 cells upregulate genes involved in cell cycle arrest and represses genes that promote cell cycle progression, proliferation and survival. Cells were transfected with DLL1-siRNA1 or Ctr-siRNA. (A) At 55–70 hours following transfection total RNA was extracted and the expression of genes that control cell cycle arrest (1), cell cycle transition (2), proliferation (3), and survival (4) were quantified by qRT-PCR. The values were normalized against the HPRT1 mRNA levels in the same sample and calculated as fold change relative to control cells transfected with Ctr-siRNA. Results are represented as mean values (+ SD) from four independent assays. (B) The expression levels of the indicated proteins were determined by immunoblotting 72 hours after transfection in total soluble protein extracts. α-tubulin was used as an internal control. The graph shows relative expression of these proteins in cells transfected with DLL1-siRNA1 as compared to Ctr-siRNA transfected cells of at least three independent experiments. *, P

    Journal: PLoS ONE

    Article Title: The Notch ligand DLL1 exerts carcinogenic features in human breast cancer cells

    doi: 10.1371/journal.pone.0217002

    Figure Lengend Snippet: DLL1 downregulation in MCF-7 cells upregulate genes involved in cell cycle arrest and represses genes that promote cell cycle progression, proliferation and survival. Cells were transfected with DLL1-siRNA1 or Ctr-siRNA. (A) At 55–70 hours following transfection total RNA was extracted and the expression of genes that control cell cycle arrest (1), cell cycle transition (2), proliferation (3), and survival (4) were quantified by qRT-PCR. The values were normalized against the HPRT1 mRNA levels in the same sample and calculated as fold change relative to control cells transfected with Ctr-siRNA. Results are represented as mean values (+ SD) from four independent assays. (B) The expression levels of the indicated proteins were determined by immunoblotting 72 hours after transfection in total soluble protein extracts. α-tubulin was used as an internal control. The graph shows relative expression of these proteins in cells transfected with DLL1-siRNA1 as compared to Ctr-siRNA transfected cells of at least three independent experiments. *, P

    Article Snippet: Construction of recombinant human DLL1 expression plasmid, protein production, and purification The cDNA fragment containing the human DLL1 signal peptide and sequences encoding the full extracellular domain was amplified by polymerase chain reaction from human DLL1 cDNA (Sino Biological, #HG11635-M, Beijing, China) using the primers: Fw: 5´- ATAGAATTCGCCGCCACCATGGGCAGTCGGTGCGCGCTGGC-3´ ; and Rev: 5´- GTCAGATCTGGATCCACGCGGAACCAGCCAGGGGAATGGCCCGCCCTG-3´ .

    Techniques: Transfection, Expressing, Quantitative RT-PCR