granulocyte macrophage colony stimulating factor  (Thermo Fisher)


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    Name:
    Granulocyte Macrophage Colony Stimulating Factor Recombinant Protein
    Description:
    Granulocyte Macrophage Colony Stimulating Factor Recombinant Protein for Western Blot ELISA Ctrl
    Catalog Number:
    sgmcsf
    Price:
    None
    Applications:
    Protein Assays and Analysis|Protein Biology
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher granulocyte macrophage colony stimulating factor
    Reverse effects of exogenous mevalonate (MEV) to the pitavastatin (PTV) inhibition. The inhibitory effects of PTV (10 nM) on lipopolysaccharide (LPS)-induced interleukin (IL)-6, IL-8 and <t>granulocyte–macrophage</t> <t>colony-stimulating</t> <t>factor</t> (GM-CSF)
    Granulocyte Macrophage Colony Stimulating Factor Recombinant Protein for Western Blot ELISA Ctrl
    https://www.bioz.com/result/granulocyte macrophage colony stimulating factor/product/Thermo Fisher
    Average 99 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    granulocyte macrophage colony stimulating factor - by Bioz Stars, 2020-11
    99/100 stars

    Images

    1) Product Images from "Inhibitory effect of statins on inflammatory cytokine production from human bronchial epithelial cells"

    Article Title: Inhibitory effect of statins on inflammatory cytokine production from human bronchial epithelial cells

    Journal: Clinical and Experimental Immunology

    doi: 10.1111/j.1365-2249.2012.04564.x

    Reverse effects of exogenous mevalonate (MEV) to the pitavastatin (PTV) inhibition. The inhibitory effects of PTV (10 nM) on lipopolysaccharide (LPS)-induced interleukin (IL)-6, IL-8 and granulocyte–macrophage colony-stimulating factor (GM-CSF)
    Figure Legend Snippet: Reverse effects of exogenous mevalonate (MEV) to the pitavastatin (PTV) inhibition. The inhibitory effects of PTV (10 nM) on lipopolysaccharide (LPS)-induced interleukin (IL)-6, IL-8 and granulocyte–macrophage colony-stimulating factor (GM-CSF)

    Techniques Used: Inhibition

    The inhibitory effects of pitavastatin (PTV) on inflammatory cytokine production. Interleukin (IL)-6, IL-8 and granulocyte–macrophage colony-stimulating factor (GM-CSF) mRNA expression in lipopolysaccharide (LPS)-stimulated BEAS-2B cells was inhibited
    Figure Legend Snippet: The inhibitory effects of pitavastatin (PTV) on inflammatory cytokine production. Interleukin (IL)-6, IL-8 and granulocyte–macrophage colony-stimulating factor (GM-CSF) mRNA expression in lipopolysaccharide (LPS)-stimulated BEAS-2B cells was inhibited

    Techniques Used: Expressing

    The effects of exogenous mevalonate on the cytokine production. Interleukin (IL)-6 and IL-8 mRNA expression, but not granulocyte–macrophage colony-stimulating factor (GM-CSF) mRNA expression, in lipopolysaccharide (LPS)-stimulated BEAS-2B cells
    Figure Legend Snippet: The effects of exogenous mevalonate on the cytokine production. Interleukin (IL)-6 and IL-8 mRNA expression, but not granulocyte–macrophage colony-stimulating factor (GM-CSF) mRNA expression, in lipopolysaccharide (LPS)-stimulated BEAS-2B cells

    Techniques Used: Expressing

    2) Product Images from "Th2 and eosinophil responses suppress inflammatory arthritis"

    Article Title: Th2 and eosinophil responses suppress inflammatory arthritis

    Journal: Nature Communications

    doi: 10.1038/ncomms11596

    N. brasiliensis infection induces Th2 response and alleviates arthritis in TNFα- mediated arthritis. ( a ) Arthritis score from 5-week-old hTNFtg mice with or without N. brasiliensis (Nb) challenge. Data are pooled from two independent experiments ( n =9 per group). ( b ) Haematoxylin/eosin (H E) and tartrate-resistant acid phosphatase (TRAP) staining of the hind paw from 9-week-old hTNFtg mice with or without Nb challenge. Scale bar, 500 μm. Quantification of inflammation area, erosion area and osteoclast number (N.Oc) per paw in the hind paws of hTNFtg mice with or without Nb challenge ( n =5–8 per group). ( c ) Frequency of CD4 + IFN-γ + (Th1), CD4 + IL-4 + (Th2) and CD4 + IL-17 + (Th17) cells in the spleen of hTNFtg mice with or without Nb challenge ( n =4–5 per group). ( d ) Serum levels of IL-4, IL-5, IL-10, IL-2, IFN-γ and granulocyte–macrophage colony-stimulating factor (GM-CSF) in hTNFtg mice with or without Nb challenge ( n =4–5 per group). ( e ) Quantitative reverse transcriptase–PCR (RT–PCR) analyses of Il4 expression in the spleen and synovial extracts from 9-week-old hTNFtg mice with or without Nb challenge ( n =4–8 per group). ( f ) Quantitative RT–PCR analyses of Il5 expression in synovial extracts of hTNFtg mice with or without Nb challenge ( n =4–8 per group). Data are expressed as mean±s.e.m. Pictures are representative of 3 independent experiments. Asterisks mark statistically significant difference (* P
    Figure Legend Snippet: N. brasiliensis infection induces Th2 response and alleviates arthritis in TNFα- mediated arthritis. ( a ) Arthritis score from 5-week-old hTNFtg mice with or without N. brasiliensis (Nb) challenge. Data are pooled from two independent experiments ( n =9 per group). ( b ) Haematoxylin/eosin (H E) and tartrate-resistant acid phosphatase (TRAP) staining of the hind paw from 9-week-old hTNFtg mice with or without Nb challenge. Scale bar, 500 μm. Quantification of inflammation area, erosion area and osteoclast number (N.Oc) per paw in the hind paws of hTNFtg mice with or without Nb challenge ( n =5–8 per group). ( c ) Frequency of CD4 + IFN-γ + (Th1), CD4 + IL-4 + (Th2) and CD4 + IL-17 + (Th17) cells in the spleen of hTNFtg mice with or without Nb challenge ( n =4–5 per group). ( d ) Serum levels of IL-4, IL-5, IL-10, IL-2, IFN-γ and granulocyte–macrophage colony-stimulating factor (GM-CSF) in hTNFtg mice with or without Nb challenge ( n =4–5 per group). ( e ) Quantitative reverse transcriptase–PCR (RT–PCR) analyses of Il4 expression in the spleen and synovial extracts from 9-week-old hTNFtg mice with or without Nb challenge ( n =4–8 per group). ( f ) Quantitative RT–PCR analyses of Il5 expression in synovial extracts of hTNFtg mice with or without Nb challenge ( n =4–8 per group). Data are expressed as mean±s.e.m. Pictures are representative of 3 independent experiments. Asterisks mark statistically significant difference (* P

    Techniques Used: Infection, Mouse Assay, Staining, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing, Quantitative RT-PCR

    3) Product Images from "Myelo-erythroid commitment after burn injury is under β-adrenergic control via MafB regulation"

    Article Title: Myelo-erythroid commitment after burn injury is under β-adrenergic control via MafB regulation

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00139.2016

    A : PBMCs were cultured in the specified growth factor cocktail for 6 days. Nonadherent cells collected from culture plates were classified as multipotent (MPP), erythroid (MEP), and nonerythroid (NE) progenitors based on a combination of cell surface antigens. First, lineage-negative (lin neg ), CD34 + , and CD38 + cells (lin neg CD34 + CD38 + ) were selected for further characterization. The lin neg CD34 + CD38 + cells comprised both MafB + and MafB neg cells, which confirms their multipotency (MPPs are the solid line in the histogram and the dashed line is Ab control). Furthermore, MPPs were subdivided according to differential expressions of CD123 (IL-3Rα) and CD45RA, such that MEPs are gated as CD123 neg CD45RA neg and N-E cells as CD123 +/− CD45RA + and verified with MafB + (N-E) and MafB neg (MEP) expressions. GMCSF, granulocyte macrophage colony-stimulating factor; SCF, stem cell factor. B : representative images of PBMC-derived MEPs and MafB-expressing cells in MPP population from standard burn care (SBC) patient (E02) at post burn day 7 (PBD7) (
    Figure Legend Snippet: A : PBMCs were cultured in the specified growth factor cocktail for 6 days. Nonadherent cells collected from culture plates were classified as multipotent (MPP), erythroid (MEP), and nonerythroid (NE) progenitors based on a combination of cell surface antigens. First, lineage-negative (lin neg ), CD34 + , and CD38 + cells (lin neg CD34 + CD38 + ) were selected for further characterization. The lin neg CD34 + CD38 + cells comprised both MafB + and MafB neg cells, which confirms their multipotency (MPPs are the solid line in the histogram and the dashed line is Ab control). Furthermore, MPPs were subdivided according to differential expressions of CD123 (IL-3Rα) and CD45RA, such that MEPs are gated as CD123 neg CD45RA neg and N-E cells as CD123 +/− CD45RA + and verified with MafB + (N-E) and MafB neg (MEP) expressions. GMCSF, granulocyte macrophage colony-stimulating factor; SCF, stem cell factor. B : representative images of PBMC-derived MEPs and MafB-expressing cells in MPP population from standard burn care (SBC) patient (E02) at post burn day 7 (PBD7) (

    Techniques Used: Cell Culture, Derivative Assay, Expressing

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Mesothelin is a target of chimeric antigen receptor T cells for treating gastric cancer
    Article Snippet: .. Cytokine release assays Enzyme-linked immune absorbance assay (ELISA) kits for IL-2, interferon-γ (IFN-γ), granzyme B, and granulocyte-macrophage colony-stimulating factor (GM-CSF) were purchased from eBioscience (San Diego, CA, USA), and all ELISAs were performed according to the manufacturer’s protocols. .. T cells were cocultured with target cells at an E:T ratio of 1:2 for 18 h. The culture supernatants were then collected and analyzed for the secretion of IL-2, IFN-γ, GM-CSF, and granzyme B using ELISA kits.

    Article Title: Evaluation of Antiangiogenic Efficacy of Emilia sonchifolia (L.) DC on Tumor-Specific Neovessel Formation by Regulating MMPs, VEGF, and Proinflammatory Cytokines
    Article Snippet: .. Highly specific quantitative sandwich enzyme-linked immunosorbent assay (ELISA) kits for mouse IL-1β, IL-6, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF) were purchased from Pierce Biotechnology (Rockford, IL). .. ELISA kits for VEGF and TIMP-1was purchased from R & D Systems (Minneapolis, MN).

    Article Title: CD4+ T Cells and IFN-γ Are Required for the Development of Pneumocystis-Associated Pulmonary Hypertension
    Article Snippet: .. Concentrations of IFN-γ, IL-13, CCL2, IL-12p40, and granulocyte-macrophage colony-stimulating factor (GM-CSF) in BALF were determined using commercial enzyme-linked immunosorbent assay kits (eBioscience). .. Total RNA was collected from the lungs of BALB-STD and IFN-γ–STD mice, as well as from immunocompetent BALB/c mice that were P. murina– infected but not depleted of CD4 and from control uninfected BALB/c mice, at day 38 after infection, using a Qiagen (Valencia, CA) RNeasy maxi kit procedure.

    Sandwich ELISA:

    Article Title: Evaluation of Antiangiogenic Efficacy of Emilia sonchifolia (L.) DC on Tumor-Specific Neovessel Formation by Regulating MMPs, VEGF, and Proinflammatory Cytokines
    Article Snippet: .. Highly specific quantitative sandwich enzyme-linked immunosorbent assay (ELISA) kits for mouse IL-1β, IL-6, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF) were purchased from Pierce Biotechnology (Rockford, IL). .. ELISA kits for VEGF and TIMP-1was purchased from R & D Systems (Minneapolis, MN).

    Real-time Polymerase Chain Reaction:

    Article Title: Inhibitory effect of statins on inflammatory cytokine production from human bronchial epithelial cells
    Article Snippet: .. Cytokine mRNA levels [interleukin (IL)-6, IL-8 and granulocyte-macrophage colony-stimulating factor (GM-CSF)] were determined using the StepOnePlusTM Real-Time PCR System (Applied Biosystems Inc., Foster City, CA, USA). .. Results were normalized against the expression of an internal control [glyceraldehyde-3-phosphate dehydrogenase (GAPDH)], and relative gene expression levels were calculated using the ΔΔCt-method.

    Multiplex Assay:

    Article Title: Th2 and eosinophil responses suppress inflammatory arthritis
    Article Snippet: .. Serum cytokine levels Mouse serum level of IL-4, IL-5, IL-10, IL-2, IFN-γ, granulocyte–macrophage colony-stimulating factor and TNF were detected by Mouse Th1/Th2 MULTIPLEX Kit FlowCytomix (eBioscience) according to the manufacturer's instructions. .. IL-1β and IL-6 level in mouse serum and IL-5 and EPX serum level in patients were analysed by ELISA kit (R & D Systems, Cloud-Clone Corp for EPX), as described in the manufacturer's instructions.

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    Thermo Fisher murine gm csf elisa kit
    Characterization of exosomes isolated from ES-D3 cells. (a) Exosomes were isolated from ES-D3 cells that were transfected with either the plasmid expressing <t>GM-CSF</t> or its empty vector. Transmission electron microscopy (TEM) imaging of the ES-D3-derived exosomes. Arrow heads indicate exosomes, Scale bar 100 nM. (b) Western blot analysis of the expression of the indicated exosomal markers, endoplasmic reticulum (ER) markers, mitochondrial markers, and cytosolic markers that are expressed either in the whole cell extracts (WCE) or in the exosomes. PDI, protein disulfide isomerase; cyto C, cytochrome C. Molecular weights markers (kD) are labeled on the left. (c) The amounts of GM-CSF in exosomes isolated from GM-CSF-expressing ES-D3 cells or from vector control ES-D3 cells were evaluated by <t>ELISA.</t> Exosomes pretreated with or without 0.05% Tween-20 were added to an ELISA plate. ELISA was carried out using washing buffer containing either PBS only or PBS + 0.05% Tween-20. The data are shown as mean ± SD of three independent ELISA experiments. ***, p
    Murine Gm Csf Elisa Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    94
    Thermo Fisher gm csf
    Characterization of exosomes isolated from <t>ES-D3</t> cells. (a) Exosomes were isolated from ES-D3 cells that were transfected with either the plasmid expressing <t>GM-CSF</t> or its empty vector. Transmission electron microscopy (TEM) imaging of the ES-D3-derived exosomes. Arrow heads indicate exosomes, Scale bar 100 nM. (b) Western blot analysis of the expression of the indicated exosomal markers, endoplasmic reticulum (ER) markers, mitochondrial markers, and cytosolic markers that are expressed either in the whole cell extracts (WCE) or in the exosomes. PDI, protein disulfide isomerase; cyto C, cytochrome C. Molecular weights markers (kD) are labeled on the left. (c) The amounts of GM-CSF in exosomes isolated from GM-CSF-expressing ES-D3 cells or from vector control ES-D3 cells were evaluated by ELISA. Exosomes pretreated with or without 0.05% Tween-20 were added to an ELISA plate. ELISA was carried out using washing buffer containing either PBS only or PBS + 0.05% Tween-20. The data are shown as mean ± SD of three independent ELISA experiments. ***, p
    Gm Csf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 191 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gm csf/product/Thermo Fisher
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    92
    Thermo Fisher csf xb buffer
    Characterization of pSer335 <t>XErp1</t> antibody <t>CSF</t> extract was treated with Myc‐XErp1 IVT carrying the indicated combinations of the mutations DSG − (S33N S38N), DSA − (S284N S288N), ZBR − (C583A) and CaMKII − (T195A). Calcium was added, samples were taken at the indicated time points and as indicated treated with λ‐phosphatase. Samples were immunoblotted for the Myc‐tag and cyclin B2. The cyclin B2 membrane was stripped and reprobed for α‐tubulin. Asterisk indicates unspecific bands. Several lanes were removed at the dashed line. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT at the indicated dilutions. An empty IVT not expressing XErp1 and an untreated condition were used as controls. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, pSer335 XErp1, and α‐tubulin. The XErp1 membrane was stripped and reprobed for the Myc‐tag. Asterisks indicate unspecific bands. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT that was either wild‐type or mutated to alanine at Ser335. An empty IVT reaction not expressing Myc‐XErp1 served as control. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, the Myc‐tag, pSer335 XErp1, and α‐tubulin. Asterisks indicate unspecific bands. Source data are available online for this figure.
    Csf Xb Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of exosomes isolated from ES-D3 cells. (a) Exosomes were isolated from ES-D3 cells that were transfected with either the plasmid expressing GM-CSF or its empty vector. Transmission electron microscopy (TEM) imaging of the ES-D3-derived exosomes. Arrow heads indicate exosomes, Scale bar 100 nM. (b) Western blot analysis of the expression of the indicated exosomal markers, endoplasmic reticulum (ER) markers, mitochondrial markers, and cytosolic markers that are expressed either in the whole cell extracts (WCE) or in the exosomes. PDI, protein disulfide isomerase; cyto C, cytochrome C. Molecular weights markers (kD) are labeled on the left. (c) The amounts of GM-CSF in exosomes isolated from GM-CSF-expressing ES-D3 cells or from vector control ES-D3 cells were evaluated by ELISA. Exosomes pretreated with or without 0.05% Tween-20 were added to an ELISA plate. ELISA was carried out using washing buffer containing either PBS only or PBS + 0.05% Tween-20. The data are shown as mean ± SD of three independent ELISA experiments. ***, p

    Journal: Oncoimmunology

    Article Title: Exosomes from GM-CSF expressing embryonic stem cells are an effective prophylactic vaccine for cancer prevention

    doi: 10.1080/2162402X.2018.1561119

    Figure Lengend Snippet: Characterization of exosomes isolated from ES-D3 cells. (a) Exosomes were isolated from ES-D3 cells that were transfected with either the plasmid expressing GM-CSF or its empty vector. Transmission electron microscopy (TEM) imaging of the ES-D3-derived exosomes. Arrow heads indicate exosomes, Scale bar 100 nM. (b) Western blot analysis of the expression of the indicated exosomal markers, endoplasmic reticulum (ER) markers, mitochondrial markers, and cytosolic markers that are expressed either in the whole cell extracts (WCE) or in the exosomes. PDI, protein disulfide isomerase; cyto C, cytochrome C. Molecular weights markers (kD) are labeled on the left. (c) The amounts of GM-CSF in exosomes isolated from GM-CSF-expressing ES-D3 cells or from vector control ES-D3 cells were evaluated by ELISA. Exosomes pretreated with or without 0.05% Tween-20 were added to an ELISA plate. ELISA was carried out using washing buffer containing either PBS only or PBS + 0.05% Tween-20. The data are shown as mean ± SD of three independent ELISA experiments. ***, p

    Article Snippet: The concentrations of GM-CSF in the ES-D3 cell supernatants were determined using a murine GM-CSF ELISA kit (88733422; Thermo Fisher) following manufacturer’s protocol.

    Techniques: Isolation, Transfection, Plasmid Preparation, Expressing, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy, Imaging, Derivative Assay, Western Blot, Labeling, Enzyme-linked Immunosorbent Assay

    Murine embryonic stem cells expressing GM-CSF maintain their pluripotency. (a) Schematic diagram of the plasmid with the EF1-α promoter driving GM-CSF expression. (b) Expression of GFP in GM-CSF-expressing ES-D3 cells and in empty vector control ES-D3 cells was evaluated by flow cytometry. (c) GM-CSF levels in transfected ES-D3 cells. ELISA measurements of GM-CSF concentrations in the medium of the indicated cells. The data are shown as mean ± standard deviations (mean ± SD) of three independements, **, p

    Journal: Oncoimmunology

    Article Title: Exosomes from GM-CSF expressing embryonic stem cells are an effective prophylactic vaccine for cancer prevention

    doi: 10.1080/2162402X.2018.1561119

    Figure Lengend Snippet: Murine embryonic stem cells expressing GM-CSF maintain their pluripotency. (a) Schematic diagram of the plasmid with the EF1-α promoter driving GM-CSF expression. (b) Expression of GFP in GM-CSF-expressing ES-D3 cells and in empty vector control ES-D3 cells was evaluated by flow cytometry. (c) GM-CSF levels in transfected ES-D3 cells. ELISA measurements of GM-CSF concentrations in the medium of the indicated cells. The data are shown as mean ± standard deviations (mean ± SD) of three independements, **, p

    Article Snippet: The concentrations of GM-CSF in the ES-D3 cell supernatants were determined using a murine GM-CSF ELISA kit (88733422; Thermo Fisher) following manufacturer’s protocol.

    Techniques: Expressing, Plasmid Preparation, Flow Cytometry, Cytometry, Transfection, Enzyme-linked Immunosorbent Assay

    Characterization of exosomes isolated from ES-D3 cells. (a) Exosomes were isolated from ES-D3 cells that were transfected with either the plasmid expressing GM-CSF or its empty vector. Transmission electron microscopy (TEM) imaging of the ES-D3-derived exosomes. Arrow heads indicate exosomes, Scale bar 100 nM. (b) Western blot analysis of the expression of the indicated exosomal markers, endoplasmic reticulum (ER) markers, mitochondrial markers, and cytosolic markers that are expressed either in the whole cell extracts (WCE) or in the exosomes. PDI, protein disulfide isomerase; cyto C, cytochrome C. Molecular weights markers (kD) are labeled on the left. (c) The amounts of GM-CSF in exosomes isolated from GM-CSF-expressing ES-D3 cells or from vector control ES-D3 cells were evaluated by ELISA. Exosomes pretreated with or without 0.05% Tween-20 were added to an ELISA plate. ELISA was carried out using washing buffer containing either PBS only or PBS + 0.05% Tween-20. The data are shown as mean ± SD of three independent ELISA experiments. ***, p

    Journal: Oncoimmunology

    Article Title: Exosomes from GM-CSF expressing embryonic stem cells are an effective prophylactic vaccine for cancer prevention

    doi: 10.1080/2162402X.2018.1561119

    Figure Lengend Snippet: Characterization of exosomes isolated from ES-D3 cells. (a) Exosomes were isolated from ES-D3 cells that were transfected with either the plasmid expressing GM-CSF or its empty vector. Transmission electron microscopy (TEM) imaging of the ES-D3-derived exosomes. Arrow heads indicate exosomes, Scale bar 100 nM. (b) Western blot analysis of the expression of the indicated exosomal markers, endoplasmic reticulum (ER) markers, mitochondrial markers, and cytosolic markers that are expressed either in the whole cell extracts (WCE) or in the exosomes. PDI, protein disulfide isomerase; cyto C, cytochrome C. Molecular weights markers (kD) are labeled on the left. (c) The amounts of GM-CSF in exosomes isolated from GM-CSF-expressing ES-D3 cells or from vector control ES-D3 cells were evaluated by ELISA. Exosomes pretreated with or without 0.05% Tween-20 were added to an ELISA plate. ELISA was carried out using washing buffer containing either PBS only or PBS + 0.05% Tween-20. The data are shown as mean ± SD of three independent ELISA experiments. ***, p

    Article Snippet: The concentrations of GM-CSF in the ES-D3 cell supernatants were determined using a murine GM-CSF ELISA kit (88733422; Thermo Fisher) following manufacturer’s protocol.

    Techniques: Isolation, Transfection, Plasmid Preparation, Expressing, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy, Imaging, Derivative Assay, Western Blot, Labeling, Enzyme-linked Immunosorbent Assay

    ESC-derived exosome vaccination decreases T regulatory (T reg ) cells and increases the ratio of effector CD8 + T cells to T reg in the tumors. C57BL/6 mice (n = 4 per group) were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes from vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank prior to s.c. challenge with LLC on day 14. Mice were euthanized 15–18 days after tumor challenge, tumors were removed, enzymatically digested, and tumor-infiltrating cells were harvested from control and vaccinated mice and analyzed by flow cytometry. (a) Dot plots showing the percentages of CD3 + CD4 + Foxp3 + T regs in CD45 + tumor-infiltrating cells obtained from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Numbers in quadrants represent the percentages of each subpopulation. (b) Bar graphs showing the percentages of CD3 + CD4 + Foxp3 + T regs sub-populations in CD45 + tumor infiltrating cells from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Results are expressed as percentages of total CD45 + cells (n = 4 per group, mean ± SD, *, p

    Journal: Oncoimmunology

    Article Title: Exosomes from GM-CSF expressing embryonic stem cells are an effective prophylactic vaccine for cancer prevention

    doi: 10.1080/2162402X.2018.1561119

    Figure Lengend Snippet: ESC-derived exosome vaccination decreases T regulatory (T reg ) cells and increases the ratio of effector CD8 + T cells to T reg in the tumors. C57BL/6 mice (n = 4 per group) were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes from vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank prior to s.c. challenge with LLC on day 14. Mice were euthanized 15–18 days after tumor challenge, tumors were removed, enzymatically digested, and tumor-infiltrating cells were harvested from control and vaccinated mice and analyzed by flow cytometry. (a) Dot plots showing the percentages of CD3 + CD4 + Foxp3 + T regs in CD45 + tumor-infiltrating cells obtained from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Numbers in quadrants represent the percentages of each subpopulation. (b) Bar graphs showing the percentages of CD3 + CD4 + Foxp3 + T regs sub-populations in CD45 + tumor infiltrating cells from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Results are expressed as percentages of total CD45 + cells (n = 4 per group, mean ± SD, *, p

    Article Snippet: The concentrations of GM-CSF in the ES-D3 cell supernatants were determined using a murine GM-CSF ELISA kit (88733422; Thermo Fisher) following manufacturer’s protocol.

    Techniques: Derivative Assay, Mouse Assay, Plasmid Preparation, Isolation, Expressing, Flow Cytometry, Cytometry

    ESC-derived exosome vaccination induces Th1-mediated cytokine responses in splenic CD8 + T cells. (a–c) C57BL/6 mice (n = 6 per group) were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes isolated from vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank. Ten days after the boost, mice were euthanized and spleens were removed. Splenocytes from vaccinated and control mice were co-cultured with LLC lysate (50 μg/mL) for an additional 4 days. Effector cells were then restimulated for 6 h with LLC lysate (50 μg/mL) in the presence of Brefeldin A (1 µL/mL of the culture medium). After restimulation, cells were harvested, Fc receptors were blocked, and stained for surface expression of CD3, CD8 and intracellular expression of cytokines and analyzed by flow cytometry. (a) Dot plots showing IFN-γ expression in CD8 + T cells in splenocyte cultures obtained from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Numbers in quadrants represent the percentages of each subpopulation. (b, c) Bar graphs showing percentages of CD8 + IFN-γ + , and CD8 + TNF-α + in splenocyte cultures derived from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Results are expressed as percentages of total cells (n = 6 per group, mean ± SD, **, p

    Journal: Oncoimmunology

    Article Title: Exosomes from GM-CSF expressing embryonic stem cells are an effective prophylactic vaccine for cancer prevention

    doi: 10.1080/2162402X.2018.1561119

    Figure Lengend Snippet: ESC-derived exosome vaccination induces Th1-mediated cytokine responses in splenic CD8 + T cells. (a–c) C57BL/6 mice (n = 6 per group) were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes isolated from vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank. Ten days after the boost, mice were euthanized and spleens were removed. Splenocytes from vaccinated and control mice were co-cultured with LLC lysate (50 μg/mL) for an additional 4 days. Effector cells were then restimulated for 6 h with LLC lysate (50 μg/mL) in the presence of Brefeldin A (1 µL/mL of the culture medium). After restimulation, cells were harvested, Fc receptors were blocked, and stained for surface expression of CD3, CD8 and intracellular expression of cytokines and analyzed by flow cytometry. (a) Dot plots showing IFN-γ expression in CD8 + T cells in splenocyte cultures obtained from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Numbers in quadrants represent the percentages of each subpopulation. (b, c) Bar graphs showing percentages of CD8 + IFN-γ + , and CD8 + TNF-α + in splenocyte cultures derived from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Results are expressed as percentages of total cells (n = 6 per group, mean ± SD, **, p

    Article Snippet: The concentrations of GM-CSF in the ES-D3 cell supernatants were determined using a murine GM-CSF ELISA kit (88733422; Thermo Fisher) following manufacturer’s protocol.

    Techniques: Derivative Assay, Mouse Assay, Isolation, Plasmid Preparation, Expressing, Cell Culture, Staining, Flow Cytometry, Cytometry

    ESC-derived exosome vaccination slows the outgrowth of an implanted mammary carcinoma. (a) Scheme of immunization. Female Balb/c mice were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes from vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank prior to s.c. challenge with syngeneic 4T1 cells on day 14. (b) Tumor growth was measured with calipers every second or third day and tumor volumes were plotted as indicated. The data represent the average tumor volumes of 20 mice/group. Error bars represent mean ± SD. (c, d) ESC-derived exosome vaccination induces Th1-mediated cytokine responses in splenic CD8 + T cells. Splenocytes from vaccinated and control mice were co-cultured with 4T1 lysate (50 μg/ml) for an additional 4 days. Effector cells were then restimulated for 6 h with 4T1 lysate (50 μg/mL) in the presence of Brefeldin A (1 µL/mL of the culture medium). After restimulation, cells were harvested, Fc receptors were blocked, and stained for surface expression of CD8 and intracellular expression of IFN-γ and analyzed by flow cytometry. (c) Dot plots showing IFN-γ expression in CD8 + cells in splenocyte cultures obtained from control, ES-exo and ES-exo/GM-CSF vaccinated mice. Numbers in quadrants represent the percentages of each subpopulation. (d) Bar graphs showing percentages of CD8 + IFN-γ + in splenocyte cultures derived from control, ES-exo and ES-exo/GM-CSF vaccinated mice. Results are expressed as percentages of total cells (n = 4 per group, mean ± SD, *, p

    Journal: Oncoimmunology

    Article Title: Exosomes from GM-CSF expressing embryonic stem cells are an effective prophylactic vaccine for cancer prevention

    doi: 10.1080/2162402X.2018.1561119

    Figure Lengend Snippet: ESC-derived exosome vaccination slows the outgrowth of an implanted mammary carcinoma. (a) Scheme of immunization. Female Balb/c mice were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes from vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank prior to s.c. challenge with syngeneic 4T1 cells on day 14. (b) Tumor growth was measured with calipers every second or third day and tumor volumes were plotted as indicated. The data represent the average tumor volumes of 20 mice/group. Error bars represent mean ± SD. (c, d) ESC-derived exosome vaccination induces Th1-mediated cytokine responses in splenic CD8 + T cells. Splenocytes from vaccinated and control mice were co-cultured with 4T1 lysate (50 μg/ml) for an additional 4 days. Effector cells were then restimulated for 6 h with 4T1 lysate (50 μg/mL) in the presence of Brefeldin A (1 µL/mL of the culture medium). After restimulation, cells were harvested, Fc receptors were blocked, and stained for surface expression of CD8 and intracellular expression of IFN-γ and analyzed by flow cytometry. (c) Dot plots showing IFN-γ expression in CD8 + cells in splenocyte cultures obtained from control, ES-exo and ES-exo/GM-CSF vaccinated mice. Numbers in quadrants represent the percentages of each subpopulation. (d) Bar graphs showing percentages of CD8 + IFN-γ + in splenocyte cultures derived from control, ES-exo and ES-exo/GM-CSF vaccinated mice. Results are expressed as percentages of total cells (n = 4 per group, mean ± SD, *, p

    Article Snippet: The concentrations of GM-CSF in the ES-D3 cell supernatants were determined using a murine GM-CSF ELISA kit (88733422; Thermo Fisher) following manufacturer’s protocol.

    Techniques: Derivative Assay, Mouse Assay, Plasmid Preparation, Isolation, Expressing, Cell Culture, Staining, Flow Cytometry, Cytometry

    Murine embryonic stem cells expressing GM-CSF maintain their pluripotency. (a) Schematic diagram of the plasmid with the EF1-α promoter driving GM-CSF expression. (b) Expression of GFP in GM-CSF-expressing ES-D3 cells and in empty vector control ES-D3 cells was evaluated by flow cytometry. (c) GM-CSF levels in transfected ES-D3 cells. ELISA measurements of GM-CSF concentrations in the medium of the indicated cells. The data are shown as mean ± standard deviations (mean ± SD) of three independements, **, p

    Journal: Oncoimmunology

    Article Title: Exosomes from GM-CSF expressing embryonic stem cells are an effective prophylactic vaccine for cancer prevention

    doi: 10.1080/2162402X.2018.1561119

    Figure Lengend Snippet: Murine embryonic stem cells expressing GM-CSF maintain their pluripotency. (a) Schematic diagram of the plasmid with the EF1-α promoter driving GM-CSF expression. (b) Expression of GFP in GM-CSF-expressing ES-D3 cells and in empty vector control ES-D3 cells was evaluated by flow cytometry. (c) GM-CSF levels in transfected ES-D3 cells. ELISA measurements of GM-CSF concentrations in the medium of the indicated cells. The data are shown as mean ± standard deviations (mean ± SD) of three independements, **, p

    Article Snippet: The concentrations of GM-CSF in the ES-D3 cell supernatants were determined using a murine GM-CSF ELISA kit (88733422; Thermo Fisher) following manufacturer’s protocol.

    Techniques: Expressing, Plasmid Preparation, Flow Cytometry, Cytometry, Transfection, Enzyme-linked Immunosorbent Assay

    ESC-derived exosome vaccination prevents the outgrowth of an implanted lung adenocarcinoma. (a) Scheme of immunization. Male C57BL/6 mice were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes isolated from the vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank prior to s.c. challenge with LLC on day 14. (b) C57BL/6 mice (20 mice/group) were immunized twice (days 0 and 7) with HBSS (control) or ES-exo or ES-exo/GM-CSF in the right flank prior to s.c. challenge with LLC on day 14. Tumor growth was monitored daily in all animals until sacrifice due to tumors exceeding 5% of body weight. The ES-exo/GM-CSF-vaccinated tumor-free mice remained so for up to 4 months later with no overt signs of distress. Results are representative of three independent experiments. ***, p

    Journal: Oncoimmunology

    Article Title: Exosomes from GM-CSF expressing embryonic stem cells are an effective prophylactic vaccine for cancer prevention

    doi: 10.1080/2162402X.2018.1561119

    Figure Lengend Snippet: ESC-derived exosome vaccination prevents the outgrowth of an implanted lung adenocarcinoma. (a) Scheme of immunization. Male C57BL/6 mice were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes isolated from the vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank prior to s.c. challenge with LLC on day 14. (b) C57BL/6 mice (20 mice/group) were immunized twice (days 0 and 7) with HBSS (control) or ES-exo or ES-exo/GM-CSF in the right flank prior to s.c. challenge with LLC on day 14. Tumor growth was monitored daily in all animals until sacrifice due to tumors exceeding 5% of body weight. The ES-exo/GM-CSF-vaccinated tumor-free mice remained so for up to 4 months later with no overt signs of distress. Results are representative of three independent experiments. ***, p

    Article Snippet: The concentrations of GM-CSF in the ES-D3 cell supernatants were determined using a murine GM-CSF ELISA kit (88733422; Thermo Fisher) following manufacturer’s protocol.

    Techniques: Derivative Assay, Mouse Assay, Isolation, Plasmid Preparation, Expressing

    ESC-derived exosome vaccination induces Th1-mediated cytokine responses in intra-tumoral CD8 + T cells. (a–c) C57BL/6 mice (n = 6 per group) were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes from vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank prior to s.c. challenge with LLC on day 14. Mice were euthanized 15–18 days after tumor challenge, tumors were removed and enzymatically digested. Tumor-infiltrating cells from vaccinated and control mice were stimulated with LLC lysate (50 μg/mL) for 24 h. Cells were then restimulated for 6 h with LLC lysate (50 μg/mL) in the presence of Brefeldin A (1 µL/mL of the culture medium). After restimulation, cells were harvested, Fc receptors were blocked, and stained for surface expression of CD8 and intracellular expression of cytokines and analyzed by flow cytometry. (a) Dot plots showing IFN-γ expression in CD45 + CD3 + CD8 + cells in tumor-infiltrating cells obtained from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Numbers in quadrants represent the percentages of each subpopulation. (b, c) Bar graphs showing percentages of CD45 + CD3 + CD8 + IFN-γ + (b) and CD45 + CD3 + CD8 + TNF-α + (c) in tumors derived from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Results are expressed as percentages of total CD45 + cells (n = 6 per group, mean ± SD, *, p

    Journal: Oncoimmunology

    Article Title: Exosomes from GM-CSF expressing embryonic stem cells are an effective prophylactic vaccine for cancer prevention

    doi: 10.1080/2162402X.2018.1561119

    Figure Lengend Snippet: ESC-derived exosome vaccination induces Th1-mediated cytokine responses in intra-tumoral CD8 + T cells. (a–c) C57BL/6 mice (n = 6 per group) were immunized twice (days 0 and 7) with vehicle only (HBSS control) or with exosomes from vector control ES-D3 cells (ES-exo) or with exosomes isolated from ES-D3 cells over-expressing GM-CSF (ES-exo/GM-CSF) in the right flank prior to s.c. challenge with LLC on day 14. Mice were euthanized 15–18 days after tumor challenge, tumors were removed and enzymatically digested. Tumor-infiltrating cells from vaccinated and control mice were stimulated with LLC lysate (50 μg/mL) for 24 h. Cells were then restimulated for 6 h with LLC lysate (50 μg/mL) in the presence of Brefeldin A (1 µL/mL of the culture medium). After restimulation, cells were harvested, Fc receptors were blocked, and stained for surface expression of CD8 and intracellular expression of cytokines and analyzed by flow cytometry. (a) Dot plots showing IFN-γ expression in CD45 + CD3 + CD8 + cells in tumor-infiltrating cells obtained from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Numbers in quadrants represent the percentages of each subpopulation. (b, c) Bar graphs showing percentages of CD45 + CD3 + CD8 + IFN-γ + (b) and CD45 + CD3 + CD8 + TNF-α + (c) in tumors derived from control, ES-exo- and ES-exo/GM-CSF-vaccinated mice. Results are expressed as percentages of total CD45 + cells (n = 6 per group, mean ± SD, *, p

    Article Snippet: The concentrations of GM-CSF in the ES-D3 cell supernatants were determined using a murine GM-CSF ELISA kit (88733422; Thermo Fisher) following manufacturer’s protocol.

    Techniques: Derivative Assay, Mouse Assay, Plasmid Preparation, Isolation, Expressing, Staining, Flow Cytometry, Cytometry

    Characterization of pSer335 XErp1 antibody CSF extract was treated with Myc‐XErp1 IVT carrying the indicated combinations of the mutations DSG − (S33N S38N), DSA − (S284N S288N), ZBR − (C583A) and CaMKII − (T195A). Calcium was added, samples were taken at the indicated time points and as indicated treated with λ‐phosphatase. Samples were immunoblotted for the Myc‐tag and cyclin B2. The cyclin B2 membrane was stripped and reprobed for α‐tubulin. Asterisk indicates unspecific bands. Several lanes were removed at the dashed line. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT at the indicated dilutions. An empty IVT not expressing XErp1 and an untreated condition were used as controls. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, pSer335 XErp1, and α‐tubulin. The XErp1 membrane was stripped and reprobed for the Myc‐tag. Asterisks indicate unspecific bands. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT that was either wild‐type or mutated to alanine at Ser335. An empty IVT reaction not expressing Myc‐XErp1 served as control. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, the Myc‐tag, pSer335 XErp1, and α‐tubulin. Asterisks indicate unspecific bands. Source data are available online for this figure.

    Journal: EMBO Reports

    Article Title: Calcineurin promotes APC/C activation at meiotic exit by acting on both XErp1 and Cdc20

    doi: 10.15252/embr.201846433

    Figure Lengend Snippet: Characterization of pSer335 XErp1 antibody CSF extract was treated with Myc‐XErp1 IVT carrying the indicated combinations of the mutations DSG − (S33N S38N), DSA − (S284N S288N), ZBR − (C583A) and CaMKII − (T195A). Calcium was added, samples were taken at the indicated time points and as indicated treated with λ‐phosphatase. Samples were immunoblotted for the Myc‐tag and cyclin B2. The cyclin B2 membrane was stripped and reprobed for α‐tubulin. Asterisk indicates unspecific bands. Several lanes were removed at the dashed line. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT at the indicated dilutions. An empty IVT not expressing XErp1 and an untreated condition were used as controls. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, pSer335 XErp1, and α‐tubulin. The XErp1 membrane was stripped and reprobed for the Myc‐tag. Asterisks indicate unspecific bands. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT that was either wild‐type or mutated to alanine at Ser335. An empty IVT reaction not expressing Myc‐XErp1 served as control. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, the Myc‐tag, pSer335 XErp1, and α‐tubulin. Asterisks indicate unspecific bands. Source data are available online for this figure.

    Article Snippet: XErp1: 2 μl Myc‐XErp1 IVT were diluted in 8 μl CSF‐XB Buffer (10 mM HEPES; 100 mM KCl; 2 mM MgCl2 ; 0.1 mM CaCl2 ; 50 mM sucrose; 5 mM EGTA; pH = 7.7) and added to 0.5 μg α‐Myc antibodies coupled to Dynabeads® Protein G (Thermo Fisher).

    Techniques: Expressing