human gm-csf carrier-free recombinant protein Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Thermo Fisher human gm csf carrier free recombinant protein
    Human Gm Csf Carrier Free Recombinant Protein, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human gm csf carrier free recombinant protein/product/Thermo Fisher
    Average 94 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    human gm csf carrier free recombinant protein - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    99
    R&D Systems carrier free recombinant human gm csf
    Carrier Free Recombinant Human Gm Csf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carrier free recombinant human gm csf/product/R&D Systems
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    carrier free recombinant human gm csf - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    94
    BioLegend carrier free mouse gm csf protein
    DZNep alters expression of MDSC master regulators. (A, B) NUP cells were incubated for 0, 1, 4 or 7 d with 20 ng/mL <t>GM-CSF,</t> 20 ng/mL IL-6 and DMSO, 1 μM RA or 0.1 μM DZNep. (A) Cebpb mRNA levels were normalized to 18s rRNA transcript level and are represented as x -fold of day 4 DMSO-treated NUP-MDSC. The corresponding WB shows C/EBPβ isoforms LAP (liver-enriched activator protein), LAP* and LIP (liver-enriched inhibitory protein) as well as <t>β-actin</t> in cell lysates of NUP-MDSC treated with DMSO, RA, or DZNep. (B) Hif1a mRNA levels were normalized to 18s rRNA transcript level and are represented as x -fold of day 4 DMSO-treated NUP-MDSC. The corresponding WB shows HIF1-α content in cell lysates of NUP-MDSC that were treated with DMSO, RA or DZNep for 4 d either in normoxia or hypoxia (20 h). Data in (A) and (B) represent mean ± SD of 3–4 independent replicates. (C) Bar diagrams are showing the amount of phosphorylated STAT3 or STAT5 for NUP-MDSC on day 0 (NUP) and day 4 (DMSO, RA, and DZNep). MFI-values represent mean ± SD of three independent replicates. (D) Bar diagrams are showing the amount of IRF8 for NUP-MDSC on day 0 (NUP) and day 4 (DMSO, RA, and DZNep). MFI-values represent mean ± SD of three independent replicates. Asterisks represent statistical significance (two-tailed t -test with Welch's correction) with ns: not significant, * p
    Carrier Free Mouse Gm Csf Protein, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carrier free mouse gm csf protein/product/BioLegend
    Average 94 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    carrier free mouse gm csf protein - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    95
    BioLegend human gm csf
    <t>GM-CSF-Producing</t> ILCs in Inflamed Joints (A) Flow cytometry analysis of GM-CSF expression by CD4 + T cells and CD4 − cells among CD45 + joint infiltrating cells in Rag2 −/− mice transferred with CD4 + T cells from WT or Csf2 −/− SKG mice. (B) Quantitative RT-PCR analysis of Csf2 , Il1b , and Tnf in CD11b + Ly-6G − (CD11b + ), CD11b + Ly-6G + (Ly-6G + ), CD4 + T cells, and ILCs sorted from arthritic joints of mannan-treated SKG mice (n = 3). mRNA expression is presented relative to the expression of Hprt1 . (C) Flow cytometry of joint infiltrating cells in Rag2 −/− mice transferred with CD4 + T cells from WT or Csf2 −/− SKG mice. Cells were stained for CD45.2 and lineage markers (a cocktail of CD3, CD4, CD8, CD11b, <t>CD11c,</t> CD19, and DX-5). (D) Proportion of ILCs in Rag2 −/− mice transferred with CD4 + T cells as shown in (C). Each symbol represents an individual mouse. Horizontal bars indicate the means. (E) Total cell number of ILCs from healthy or inflamed joints of SKG mice (n = 3). (F) Flow cytometry of synovial ILCs (CD45.2 + lineage markers-negative Thy1.2 + cells as shown in C) for Ki-67 expression. (G) Flow cytometry of synovial ILCs (CD45.2 + lineage markers-negative Thy1.2 + cells as shown in C) for cell surface expression of IL-7Ra, CD25, CCR6, c-kit, IL-33Ra, CD44, and MHC2. (H) Flow cytometry of synovial ILCs (as shown in C) for intranuclear expression of the transcription factor T-bet, Gata-3, Rorγt, and Foxp3. (I) Proportion of the transcription factor-expressing synovial ILCs (n = 3) as shown in (H). (J) Flow cytometry of synovial ILCs (as shown in C) for the expression of GM-CSF, Gata-3, and IL-13. (K) Total cell numbers of ILCs from healthy or inflamed joints of C57/BL6 (B6) mice with collagen antibody-induced arthritis (n = 3). Data are representative of two independent experiments. (L) Flow cytometry of synovial ILCs for the expression of GM-CSF and FP635 in arthritic Il17a Cre R26R FP635 SKG mice. (M) Quantitative RT-PCR analysis of Csf2 and Bhlhe40 in splenic naive CD25 − CD44 lo CD4 + T cells (naive CD4 + T) and synovial ILCs (n = 3) as shown in (C). (N) The effects of ILC depletion on arthritis development. CD4 + T cells (1 × 10 6 ) from Thy1.1 + SKG mice were adoptively transferred into Thy1.2 + Rag2 −/− mice, which were i.v. injected with 500 μg anti-Thy1.2 mAb or control Rat IgG every week (n = 19 each). The severity of arthritis was monitored every week. ∗ p
    Human Gm Csf, supplied by BioLegend, used in various techniques. Bioz Stars score: 95/100, based on 189 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human gm csf/product/BioLegend
    Average 95 stars, based on 189 article reviews
    Price from $9.99 to $1999.99
    human gm csf - by Bioz Stars, 2020-07
    95/100 stars
      Buy from Supplier

    94
    BioLegend recombinant mouse csf 2
    <t>CSF-2</t> induces AM proliferation and rescues aberrant pulmonary surfactant PC accumulation in CD44 -/- mice. (A) Representative flow cytometry plots showing the proportion of BAL AMs from CD44 -/- mice after 7 days of PBS or CSF-2 treatment. (B) Number of AMs in the BAL from CD44 -/- mice after PBS or CSF-2 treatment. (C) BAL PC concentration from CD44 -/- mice after PBS or CSF-2 treatment. (D) Representative flow cytometry histograms comparing the phenotype of AMs from CD44 -/- mice after 7 days of PBS or CSF-2 treatment. (E) Graphs comparing the MFI of SSC-A, BODIPY, CD36, and CD11c between AMs from CD44 -/- mice after 7 days of PBS or CSF-2 treatment. Data show an average of two experiments ± SD, each with three to five mice. Background MFI of autofluorescence from unlabeled CD44 +/+ and CD44 -/- cells were respectively subtracted in the flow cytometry analysis. Significance indicated as * p
    Recombinant Mouse Csf 2, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant mouse csf 2/product/BioLegend
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant mouse csf 2 - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    93
    R&D Systems mouse recombinant carrier free gm csf
    Role of TGF-β in the development of microglia in vitro. ( a and b ) MG400 analysis of (a) upregulated microglial genes and (b) downregulated microglial genes in cultured adult microglia in the presence of <t>MCSF</t> and TGF-β1 in comparison to adult microglia cultured in the presence of <t>GM-CSF</t> or MCSF alone (see Source data – Figure 6). One representative of three individual experiments is shown. (c) qPCR analysis of 6 selected microglial genes in cultured adult microglia in the presence of MCSF and TGF-β1. Gene expression level was normalized against Gapdh using ΔCt (n = 6, each biological triplicate consisted of two wells per treatment). Bars show mean normalized intensity ± s.e.m. * P
    Mouse Recombinant Carrier Free Gm Csf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse recombinant carrier free gm csf/product/R&D Systems
    Average 93 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    mouse recombinant carrier free gm csf - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    99
    R&D Systems recombinant murine gm csf
    Role of TGF-β in the development of microglia in vitro. ( a and b ) MG400 analysis of (a) upregulated microglial genes and (b) downregulated microglial genes in cultured adult microglia in the presence of <t>MCSF</t> and TGF-β1 in comparison to adult microglia cultured in the presence of <t>GM-CSF</t> or MCSF alone (see Source data – Figure 6). One representative of three individual experiments is shown. (c) qPCR analysis of 6 selected microglial genes in cultured adult microglia in the presence of MCSF and TGF-β1. Gene expression level was normalized against Gapdh using ΔCt (n = 6, each biological triplicate consisted of two wells per treatment). Bars show mean normalized intensity ± s.e.m. * P
    Recombinant Murine Gm Csf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 300 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant murine gm csf/product/R&D Systems
    Average 99 stars, based on 300 article reviews
    Price from $9.99 to $1999.99
    recombinant murine gm csf - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    DZNep alters expression of MDSC master regulators. (A, B) NUP cells were incubated for 0, 1, 4 or 7 d with 20 ng/mL GM-CSF, 20 ng/mL IL-6 and DMSO, 1 μM RA or 0.1 μM DZNep. (A) Cebpb mRNA levels were normalized to 18s rRNA transcript level and are represented as x -fold of day 4 DMSO-treated NUP-MDSC. The corresponding WB shows C/EBPβ isoforms LAP (liver-enriched activator protein), LAP* and LIP (liver-enriched inhibitory protein) as well as β-actin in cell lysates of NUP-MDSC treated with DMSO, RA, or DZNep. (B) Hif1a mRNA levels were normalized to 18s rRNA transcript level and are represented as x -fold of day 4 DMSO-treated NUP-MDSC. The corresponding WB shows HIF1-α content in cell lysates of NUP-MDSC that were treated with DMSO, RA or DZNep for 4 d either in normoxia or hypoxia (20 h). Data in (A) and (B) represent mean ± SD of 3–4 independent replicates. (C) Bar diagrams are showing the amount of phosphorylated STAT3 or STAT5 for NUP-MDSC on day 0 (NUP) and day 4 (DMSO, RA, and DZNep). MFI-values represent mean ± SD of three independent replicates. (D) Bar diagrams are showing the amount of IRF8 for NUP-MDSC on day 0 (NUP) and day 4 (DMSO, RA, and DZNep). MFI-values represent mean ± SD of three independent replicates. Asterisks represent statistical significance (two-tailed t -test with Welch's correction) with ns: not significant, * p

    Journal: Oncoimmunology

    Article Title: Identification of inhibitors of myeloid-derived suppressor cells activity through phenotypic chemical screening

    doi: 10.1080/2162402X.2016.1258503

    Figure Lengend Snippet: DZNep alters expression of MDSC master regulators. (A, B) NUP cells were incubated for 0, 1, 4 or 7 d with 20 ng/mL GM-CSF, 20 ng/mL IL-6 and DMSO, 1 μM RA or 0.1 μM DZNep. (A) Cebpb mRNA levels were normalized to 18s rRNA transcript level and are represented as x -fold of day 4 DMSO-treated NUP-MDSC. The corresponding WB shows C/EBPβ isoforms LAP (liver-enriched activator protein), LAP* and LIP (liver-enriched inhibitory protein) as well as β-actin in cell lysates of NUP-MDSC treated with DMSO, RA, or DZNep. (B) Hif1a mRNA levels were normalized to 18s rRNA transcript level and are represented as x -fold of day 4 DMSO-treated NUP-MDSC. The corresponding WB shows HIF1-α content in cell lysates of NUP-MDSC that were treated with DMSO, RA or DZNep for 4 d either in normoxia or hypoxia (20 h). Data in (A) and (B) represent mean ± SD of 3–4 independent replicates. (C) Bar diagrams are showing the amount of phosphorylated STAT3 or STAT5 for NUP-MDSC on day 0 (NUP) and day 4 (DMSO, RA, and DZNep). MFI-values represent mean ± SD of three independent replicates. (D) Bar diagrams are showing the amount of IRF8 for NUP-MDSC on day 0 (NUP) and day 4 (DMSO, RA, and DZNep). MFI-values represent mean ± SD of three independent replicates. Asterisks represent statistical significance (two-tailed t -test with Welch's correction) with ns: not significant, * p

    Article Snippet: To induce MDSC differentiation, the NUP cells (not older than 4 weeks) were pelleted and resuspended in RPMI1640 medium (10% FBS, 100U/mL penicillin, 100µg/mL streptomycin, 1 mM sodium pyruvate, 2 mM β-mercaptoethanol, 1X non-essential amino acids) containing 20 ng/mL IL-6 and 20 ng/mL GM-CSF (BioLegend, #576304).

    Techniques: Expressing, Incubation, Western Blot, Two Tailed Test

    GM-CSF-Producing ILCs in Inflamed Joints (A) Flow cytometry analysis of GM-CSF expression by CD4 + T cells and CD4 − cells among CD45 + joint infiltrating cells in Rag2 −/− mice transferred with CD4 + T cells from WT or Csf2 −/− SKG mice. (B) Quantitative RT-PCR analysis of Csf2 , Il1b , and Tnf in CD11b + Ly-6G − (CD11b + ), CD11b + Ly-6G + (Ly-6G + ), CD4 + T cells, and ILCs sorted from arthritic joints of mannan-treated SKG mice (n = 3). mRNA expression is presented relative to the expression of Hprt1 . (C) Flow cytometry of joint infiltrating cells in Rag2 −/− mice transferred with CD4 + T cells from WT or Csf2 −/− SKG mice. Cells were stained for CD45.2 and lineage markers (a cocktail of CD3, CD4, CD8, CD11b, CD11c, CD19, and DX-5). (D) Proportion of ILCs in Rag2 −/− mice transferred with CD4 + T cells as shown in (C). Each symbol represents an individual mouse. Horizontal bars indicate the means. (E) Total cell number of ILCs from healthy or inflamed joints of SKG mice (n = 3). (F) Flow cytometry of synovial ILCs (CD45.2 + lineage markers-negative Thy1.2 + cells as shown in C) for Ki-67 expression. (G) Flow cytometry of synovial ILCs (CD45.2 + lineage markers-negative Thy1.2 + cells as shown in C) for cell surface expression of IL-7Ra, CD25, CCR6, c-kit, IL-33Ra, CD44, and MHC2. (H) Flow cytometry of synovial ILCs (as shown in C) for intranuclear expression of the transcription factor T-bet, Gata-3, Rorγt, and Foxp3. (I) Proportion of the transcription factor-expressing synovial ILCs (n = 3) as shown in (H). (J) Flow cytometry of synovial ILCs (as shown in C) for the expression of GM-CSF, Gata-3, and IL-13. (K) Total cell numbers of ILCs from healthy or inflamed joints of C57/BL6 (B6) mice with collagen antibody-induced arthritis (n = 3). Data are representative of two independent experiments. (L) Flow cytometry of synovial ILCs for the expression of GM-CSF and FP635 in arthritic Il17a Cre R26R FP635 SKG mice. (M) Quantitative RT-PCR analysis of Csf2 and Bhlhe40 in splenic naive CD25 − CD44 lo CD4 + T cells (naive CD4 + T) and synovial ILCs (n = 3) as shown in (C). (N) The effects of ILC depletion on arthritis development. CD4 + T cells (1 × 10 6 ) from Thy1.1 + SKG mice were adoptively transferred into Thy1.2 + Rag2 −/− mice, which were i.v. injected with 500 μg anti-Thy1.2 mAb or control Rat IgG every week (n = 19 each). The severity of arthritis was monitored every week. ∗ p

    Journal: Immunity

    Article Title: Autoimmune Th17 Cells Induced Synovial Stromal and Innate Lymphoid Cell Secretion of the Cytokine GM-CSF to Initiate and Augment Autoimmune Arthritis

    doi: 10.1016/j.immuni.2018.04.009

    Figure Lengend Snippet: GM-CSF-Producing ILCs in Inflamed Joints (A) Flow cytometry analysis of GM-CSF expression by CD4 + T cells and CD4 − cells among CD45 + joint infiltrating cells in Rag2 −/− mice transferred with CD4 + T cells from WT or Csf2 −/− SKG mice. (B) Quantitative RT-PCR analysis of Csf2 , Il1b , and Tnf in CD11b + Ly-6G − (CD11b + ), CD11b + Ly-6G + (Ly-6G + ), CD4 + T cells, and ILCs sorted from arthritic joints of mannan-treated SKG mice (n = 3). mRNA expression is presented relative to the expression of Hprt1 . (C) Flow cytometry of joint infiltrating cells in Rag2 −/− mice transferred with CD4 + T cells from WT or Csf2 −/− SKG mice. Cells were stained for CD45.2 and lineage markers (a cocktail of CD3, CD4, CD8, CD11b, CD11c, CD19, and DX-5). (D) Proportion of ILCs in Rag2 −/− mice transferred with CD4 + T cells as shown in (C). Each symbol represents an individual mouse. Horizontal bars indicate the means. (E) Total cell number of ILCs from healthy or inflamed joints of SKG mice (n = 3). (F) Flow cytometry of synovial ILCs (CD45.2 + lineage markers-negative Thy1.2 + cells as shown in C) for Ki-67 expression. (G) Flow cytometry of synovial ILCs (CD45.2 + lineage markers-negative Thy1.2 + cells as shown in C) for cell surface expression of IL-7Ra, CD25, CCR6, c-kit, IL-33Ra, CD44, and MHC2. (H) Flow cytometry of synovial ILCs (as shown in C) for intranuclear expression of the transcription factor T-bet, Gata-3, Rorγt, and Foxp3. (I) Proportion of the transcription factor-expressing synovial ILCs (n = 3) as shown in (H). (J) Flow cytometry of synovial ILCs (as shown in C) for the expression of GM-CSF, Gata-3, and IL-13. (K) Total cell numbers of ILCs from healthy or inflamed joints of C57/BL6 (B6) mice with collagen antibody-induced arthritis (n = 3). Data are representative of two independent experiments. (L) Flow cytometry of synovial ILCs for the expression of GM-CSF and FP635 in arthritic Il17a Cre R26R FP635 SKG mice. (M) Quantitative RT-PCR analysis of Csf2 and Bhlhe40 in splenic naive CD25 − CD44 lo CD4 + T cells (naive CD4 + T) and synovial ILCs (n = 3) as shown in (C). (N) The effects of ILC depletion on arthritis development. CD4 + T cells (1 × 10 6 ) from Thy1.1 + SKG mice were adoptively transferred into Thy1.2 + Rag2 −/− mice, which were i.v. injected with 500 μg anti-Thy1.2 mAb or control Rat IgG every week (n = 19 each). The severity of arthritis was monitored every week. ∗ p

    Article Snippet: The following monoclonal antibodies were used for flow cytometry analysis (BD FACSCantoII) and cell sorting (BD FACSAria SORP): anti-mouse CCR6 (29-2L17, Biolegend), CD3e (145-2C11, BD Biosciences), CD4 (RM4-4, Biolegend), CD8 (53-6.7, Biolegend), CD11b (M1/70, Biolegend), CD11c (HL3, BD Biosciences), CD16/32 (2.4G2, BD Biosciences), CD19 (1D3, BD Biosciences), CD25 (PC61, BD Biosciences), CD44 (IM7, BD Biosciences), CD45.2 (104, Biolegend), c-Kit (2B8, Biolegend), Foxp3 (FJK-16S, eBioscience), Gata-3 (TWAI, eBioscience), GM-CSF (MP1-22E9, BD Biosciences), GM-CSFRa (698423, R & D systems), IFN-γ (XMG1.2, eBioscience), IL-13 (eBio13A, eBioscience), IL-17 (TC11-18H10.1, Biolegend), IL-7Ra (SB/199, BD Biosciences), Ki-67 (MKI67, BD Biosciences), IL-33Ra (D1H9, Biolegend), Ly-6C (AL-21, BD Biosciences), Ly-6G (1A8, Biologend), MHC2 (M5/114.15.2, Biolegend), Pan-NK (DX-5, eBioscience), Podoplanin (8.1.1, Biolegend), Rorγt (AFKJS-9, eBioscience), TCR-β (H57-597, Biolegend), T-bet (4B10, Biolegend), Thy1.1 (OX-7, BD Biosciences), Thy1.2 (53-2.1, Biolegend), anti-human CD3 (UCHT1, Biolegend), CD4 (OKT4, Biolegend), CD8 (SK1, Biolegend), CD11b (M1/70, eBioscience), CD11c (3.9, Biolegend), CD19 (HIB19, Biolegend), CD45 (HI30, Biolegend), CD56 (HCD56, Biolegend), GM-CSF (BVD2-21C11, Biolegend), IFN-γ (4S.B3, Biolegend), IL-13 (JES10-5A2, Biolegend), IL-17 (BL168, Biolegend), PECy7-Streptavidin (BD Biosciences).

    Techniques: Flow Cytometry, Cytometry, Expressing, Mouse Assay, Quantitative RT-PCR, Staining, Injection

    GM-CSF from Non-T Cells Is Crucial for the Initiation of Autoimmune Arthritis (A) Experimental design of adoptive transfer of CD4 + T cells from WT or Csf2 −/− SKG mice into Rag2 −/− or Csf2 −/− Rag2 −/− mice. Arthritis scores of four groups (a–d) of mice were assessed 3 months after transfer of 1 × 10 6 CD4 + T cells. (B) Arthritis scores of the four groups mice (n = 15 or 16 each) shown in (A). Horizontal bars indicate the means. (C) Representative joint histology of the groups shown in (A). Scale bars indicate 200 μm. (D) Flow cytometry of splenic CD4 + T cells stained for intracellular IL-17 and GM-CSF or IFN-γ. (E) Proportion of IL-17-producing CD4 + T cells from individual mice as shown in (D). Vertical bars mean SD (n = 3). ∗∗ p

    Journal: Immunity

    Article Title: Autoimmune Th17 Cells Induced Synovial Stromal and Innate Lymphoid Cell Secretion of the Cytokine GM-CSF to Initiate and Augment Autoimmune Arthritis

    doi: 10.1016/j.immuni.2018.04.009

    Figure Lengend Snippet: GM-CSF from Non-T Cells Is Crucial for the Initiation of Autoimmune Arthritis (A) Experimental design of adoptive transfer of CD4 + T cells from WT or Csf2 −/− SKG mice into Rag2 −/− or Csf2 −/− Rag2 −/− mice. Arthritis scores of four groups (a–d) of mice were assessed 3 months after transfer of 1 × 10 6 CD4 + T cells. (B) Arthritis scores of the four groups mice (n = 15 or 16 each) shown in (A). Horizontal bars indicate the means. (C) Representative joint histology of the groups shown in (A). Scale bars indicate 200 μm. (D) Flow cytometry of splenic CD4 + T cells stained for intracellular IL-17 and GM-CSF or IFN-γ. (E) Proportion of IL-17-producing CD4 + T cells from individual mice as shown in (D). Vertical bars mean SD (n = 3). ∗∗ p

    Article Snippet: The following monoclonal antibodies were used for flow cytometry analysis (BD FACSCantoII) and cell sorting (BD FACSAria SORP): anti-mouse CCR6 (29-2L17, Biolegend), CD3e (145-2C11, BD Biosciences), CD4 (RM4-4, Biolegend), CD8 (53-6.7, Biolegend), CD11b (M1/70, Biolegend), CD11c (HL3, BD Biosciences), CD16/32 (2.4G2, BD Biosciences), CD19 (1D3, BD Biosciences), CD25 (PC61, BD Biosciences), CD44 (IM7, BD Biosciences), CD45.2 (104, Biolegend), c-Kit (2B8, Biolegend), Foxp3 (FJK-16S, eBioscience), Gata-3 (TWAI, eBioscience), GM-CSF (MP1-22E9, BD Biosciences), GM-CSFRa (698423, R & D systems), IFN-γ (XMG1.2, eBioscience), IL-13 (eBio13A, eBioscience), IL-17 (TC11-18H10.1, Biolegend), IL-7Ra (SB/199, BD Biosciences), Ki-67 (MKI67, BD Biosciences), IL-33Ra (D1H9, Biolegend), Ly-6C (AL-21, BD Biosciences), Ly-6G (1A8, Biologend), MHC2 (M5/114.15.2, Biolegend), Pan-NK (DX-5, eBioscience), Podoplanin (8.1.1, Biolegend), Rorγt (AFKJS-9, eBioscience), TCR-β (H57-597, Biolegend), T-bet (4B10, Biolegend), Thy1.1 (OX-7, BD Biosciences), Thy1.2 (53-2.1, Biolegend), anti-human CD3 (UCHT1, Biolegend), CD4 (OKT4, Biolegend), CD8 (SK1, Biolegend), CD11b (M1/70, eBioscience), CD11c (3.9, Biolegend), CD19 (HIB19, Biolegend), CD45 (HI30, Biolegend), CD56 (HCD56, Biolegend), GM-CSF (BVD2-21C11, Biolegend), IFN-γ (4S.B3, Biolegend), IL-13 (JES10-5A2, Biolegend), IL-17 (BL168, Biolegend), PECy7-Streptavidin (BD Biosciences).

    Techniques: Adoptive Transfer Assay, Mouse Assay, Flow Cytometry, Cytometry, Staining

    GM-CSF-Producing ILCs in Synovial Fluid of RA Patients (A) The presence of ILCs (defined as CD45 + CD3 − CD4 − CD8 − CD11b − CD11c − CD19 − CD56 − ) from peripheral blood (PB) or synovial fluid (SF) of a patient with RA or OA (left). The percentages of ILCs in PB and SF from individual RA (n = 13) or OA (n = 6) patients. The lines indicate the sample pairs of the same patients (right). (B) Total numbers of ILCs in 1 mL of SF from OA and RA patients (n = 6). Vertical bars indicate SD. (C) Flow cytometry analysis of IFN-γ, IL-13, IL-17, and GM-CSF expression by ILCs (gated as in A) in PB or SF of a RA patient (top). The percentages of cytokine-producing ILCs from individual RA patients (n = 11) (bottom). (D) Gating strategies for GM-CSF + CD45 + lineage markers-negative (ILCs), GM-CSF + CD45 + CD3 − CD11b + (myeloid cells), and GM-CSF + CD45 + CD11b − CD3 + cells (T cells). (E) Proportion of GM-CSF-producing cells (n = 3). Vertical bars indicate SD. Symbols represent individual samples. Horizontal bars indicate the means. ∗ p

    Journal: Immunity

    Article Title: Autoimmune Th17 Cells Induced Synovial Stromal and Innate Lymphoid Cell Secretion of the Cytokine GM-CSF to Initiate and Augment Autoimmune Arthritis

    doi: 10.1016/j.immuni.2018.04.009

    Figure Lengend Snippet: GM-CSF-Producing ILCs in Synovial Fluid of RA Patients (A) The presence of ILCs (defined as CD45 + CD3 − CD4 − CD8 − CD11b − CD11c − CD19 − CD56 − ) from peripheral blood (PB) or synovial fluid (SF) of a patient with RA or OA (left). The percentages of ILCs in PB and SF from individual RA (n = 13) or OA (n = 6) patients. The lines indicate the sample pairs of the same patients (right). (B) Total numbers of ILCs in 1 mL of SF from OA and RA patients (n = 6). Vertical bars indicate SD. (C) Flow cytometry analysis of IFN-γ, IL-13, IL-17, and GM-CSF expression by ILCs (gated as in A) in PB or SF of a RA patient (top). The percentages of cytokine-producing ILCs from individual RA patients (n = 11) (bottom). (D) Gating strategies for GM-CSF + CD45 + lineage markers-negative (ILCs), GM-CSF + CD45 + CD3 − CD11b + (myeloid cells), and GM-CSF + CD45 + CD11b − CD3 + cells (T cells). (E) Proportion of GM-CSF-producing cells (n = 3). Vertical bars indicate SD. Symbols represent individual samples. Horizontal bars indicate the means. ∗ p

    Article Snippet: The following monoclonal antibodies were used for flow cytometry analysis (BD FACSCantoII) and cell sorting (BD FACSAria SORP): anti-mouse CCR6 (29-2L17, Biolegend), CD3e (145-2C11, BD Biosciences), CD4 (RM4-4, Biolegend), CD8 (53-6.7, Biolegend), CD11b (M1/70, Biolegend), CD11c (HL3, BD Biosciences), CD16/32 (2.4G2, BD Biosciences), CD19 (1D3, BD Biosciences), CD25 (PC61, BD Biosciences), CD44 (IM7, BD Biosciences), CD45.2 (104, Biolegend), c-Kit (2B8, Biolegend), Foxp3 (FJK-16S, eBioscience), Gata-3 (TWAI, eBioscience), GM-CSF (MP1-22E9, BD Biosciences), GM-CSFRa (698423, R & D systems), IFN-γ (XMG1.2, eBioscience), IL-13 (eBio13A, eBioscience), IL-17 (TC11-18H10.1, Biolegend), IL-7Ra (SB/199, BD Biosciences), Ki-67 (MKI67, BD Biosciences), IL-33Ra (D1H9, Biolegend), Ly-6C (AL-21, BD Biosciences), Ly-6G (1A8, Biologend), MHC2 (M5/114.15.2, Biolegend), Pan-NK (DX-5, eBioscience), Podoplanin (8.1.1, Biolegend), Rorγt (AFKJS-9, eBioscience), TCR-β (H57-597, Biolegend), T-bet (4B10, Biolegend), Thy1.1 (OX-7, BD Biosciences), Thy1.2 (53-2.1, Biolegend), anti-human CD3 (UCHT1, Biolegend), CD4 (OKT4, Biolegend), CD8 (SK1, Biolegend), CD11b (M1/70, eBioscience), CD11c (3.9, Biolegend), CD19 (HIB19, Biolegend), CD45 (HI30, Biolegend), CD56 (HCD56, Biolegend), GM-CSF (BVD2-21C11, Biolegend), IFN-γ (4S.B3, Biolegend), IL-13 (JES10-5A2, Biolegend), IL-17 (BL168, Biolegend), PECy7-Streptavidin (BD Biosciences).

    Techniques: Flow Cytometry, Cytometry, Expressing

    Induction of GM-CSF in FLSs Stimulated with IL-17 (A) Quantitative RT-PCR analysis for the expression of designated genes in IL-17-stimulated FLSs. FLSs (2.5 × 10 4 ) were stimulated with 50 ng/mL rmIL-17 and harvested at the indicated time points. mRNA expression is presented relative to the expression of Hprt1 . (B) Quantitative RT-PCR analysis for the expression of designated genes in synoviocytes from Rag2 −/− mice with CD4 + T cell transfer. CD45 − Podoplanin + synoviocytes (3 × 10 4 ) were sorted from inflamed joints of Rag2 −/− mice 4 weeks after transfer of 1 × 10 6 SKG or Il17a −/− SKG CD4 + T cells. Vertical bars mean SD (n = 3). Data are representative of two independent experiments.

    Journal: Immunity

    Article Title: Autoimmune Th17 Cells Induced Synovial Stromal and Innate Lymphoid Cell Secretion of the Cytokine GM-CSF to Initiate and Augment Autoimmune Arthritis

    doi: 10.1016/j.immuni.2018.04.009

    Figure Lengend Snippet: Induction of GM-CSF in FLSs Stimulated with IL-17 (A) Quantitative RT-PCR analysis for the expression of designated genes in IL-17-stimulated FLSs. FLSs (2.5 × 10 4 ) were stimulated with 50 ng/mL rmIL-17 and harvested at the indicated time points. mRNA expression is presented relative to the expression of Hprt1 . (B) Quantitative RT-PCR analysis for the expression of designated genes in synoviocytes from Rag2 −/− mice with CD4 + T cell transfer. CD45 − Podoplanin + synoviocytes (3 × 10 4 ) were sorted from inflamed joints of Rag2 −/− mice 4 weeks after transfer of 1 × 10 6 SKG or Il17a −/− SKG CD4 + T cells. Vertical bars mean SD (n = 3). Data are representative of two independent experiments.

    Article Snippet: The following monoclonal antibodies were used for flow cytometry analysis (BD FACSCantoII) and cell sorting (BD FACSAria SORP): anti-mouse CCR6 (29-2L17, Biolegend), CD3e (145-2C11, BD Biosciences), CD4 (RM4-4, Biolegend), CD8 (53-6.7, Biolegend), CD11b (M1/70, Biolegend), CD11c (HL3, BD Biosciences), CD16/32 (2.4G2, BD Biosciences), CD19 (1D3, BD Biosciences), CD25 (PC61, BD Biosciences), CD44 (IM7, BD Biosciences), CD45.2 (104, Biolegend), c-Kit (2B8, Biolegend), Foxp3 (FJK-16S, eBioscience), Gata-3 (TWAI, eBioscience), GM-CSF (MP1-22E9, BD Biosciences), GM-CSFRa (698423, R & D systems), IFN-γ (XMG1.2, eBioscience), IL-13 (eBio13A, eBioscience), IL-17 (TC11-18H10.1, Biolegend), IL-7Ra (SB/199, BD Biosciences), Ki-67 (MKI67, BD Biosciences), IL-33Ra (D1H9, Biolegend), Ly-6C (AL-21, BD Biosciences), Ly-6G (1A8, Biologend), MHC2 (M5/114.15.2, Biolegend), Pan-NK (DX-5, eBioscience), Podoplanin (8.1.1, Biolegend), Rorγt (AFKJS-9, eBioscience), TCR-β (H57-597, Biolegend), T-bet (4B10, Biolegend), Thy1.1 (OX-7, BD Biosciences), Thy1.2 (53-2.1, Biolegend), anti-human CD3 (UCHT1, Biolegend), CD4 (OKT4, Biolegend), CD8 (SK1, Biolegend), CD11b (M1/70, eBioscience), CD11c (3.9, Biolegend), CD19 (HIB19, Biolegend), CD45 (HI30, Biolegend), CD56 (HCD56, Biolegend), GM-CSF (BVD2-21C11, Biolegend), IFN-γ (4S.B3, Biolegend), IL-13 (JES10-5A2, Biolegend), IL-17 (BL168, Biolegend), PECy7-Streptavidin (BD Biosciences).

    Techniques: Quantitative RT-PCR, Expressing, Mouse Assay

    GM-CSF-Producing T Helper Cells Are Dispensable for GM-CSF-Dependent Autoimmune Arthritis Development (A) Intracellular IL-17, IFN-γ, and GM-CSF staining of CD4 + T cells from popliteal LNs or inflamed joints. (B) Proportion of cytokine-producing cells in CD4 + T cells from individual mice as shown in (A). Vertical bars mean SD (n = 3). (C) Arthritis scores assessed in individual SKG, Csf2 −/− SKG, or Il17a −/− SKG mice (n = 20 each) 3 months after single i.p. injection of 20 mg mannan. (D) Intracellular IL-17, IFN-γ, and GM-CSF staining of CD4 + T cells from inflamed joints of Il17a Cre R26R eYFP SKG mice. (E) Arthritis development after adoptive transfer of CD4 + T cells from WT or Csf2 −/− SKG mice into Rag2 −/− mice (n = 17 each, SEM). The severity of arthritis was monitored every week after transfer of 1 × 10 6 CD4 + T cells. (F) Intracellular IL-17 and GM-CSF staining of CD4 + T cells from spleens and inflamed joints of Rag2 −/− mice with CD4 + T cells transfer as shown in (E). ∗ p

    Journal: Immunity

    Article Title: Autoimmune Th17 Cells Induced Synovial Stromal and Innate Lymphoid Cell Secretion of the Cytokine GM-CSF to Initiate and Augment Autoimmune Arthritis

    doi: 10.1016/j.immuni.2018.04.009

    Figure Lengend Snippet: GM-CSF-Producing T Helper Cells Are Dispensable for GM-CSF-Dependent Autoimmune Arthritis Development (A) Intracellular IL-17, IFN-γ, and GM-CSF staining of CD4 + T cells from popliteal LNs or inflamed joints. (B) Proportion of cytokine-producing cells in CD4 + T cells from individual mice as shown in (A). Vertical bars mean SD (n = 3). (C) Arthritis scores assessed in individual SKG, Csf2 −/− SKG, or Il17a −/− SKG mice (n = 20 each) 3 months after single i.p. injection of 20 mg mannan. (D) Intracellular IL-17, IFN-γ, and GM-CSF staining of CD4 + T cells from inflamed joints of Il17a Cre R26R eYFP SKG mice. (E) Arthritis development after adoptive transfer of CD4 + T cells from WT or Csf2 −/− SKG mice into Rag2 −/− mice (n = 17 each, SEM). The severity of arthritis was monitored every week after transfer of 1 × 10 6 CD4 + T cells. (F) Intracellular IL-17 and GM-CSF staining of CD4 + T cells from spleens and inflamed joints of Rag2 −/− mice with CD4 + T cells transfer as shown in (E). ∗ p

    Article Snippet: The following monoclonal antibodies were used for flow cytometry analysis (BD FACSCantoII) and cell sorting (BD FACSAria SORP): anti-mouse CCR6 (29-2L17, Biolegend), CD3e (145-2C11, BD Biosciences), CD4 (RM4-4, Biolegend), CD8 (53-6.7, Biolegend), CD11b (M1/70, Biolegend), CD11c (HL3, BD Biosciences), CD16/32 (2.4G2, BD Biosciences), CD19 (1D3, BD Biosciences), CD25 (PC61, BD Biosciences), CD44 (IM7, BD Biosciences), CD45.2 (104, Biolegend), c-Kit (2B8, Biolegend), Foxp3 (FJK-16S, eBioscience), Gata-3 (TWAI, eBioscience), GM-CSF (MP1-22E9, BD Biosciences), GM-CSFRa (698423, R & D systems), IFN-γ (XMG1.2, eBioscience), IL-13 (eBio13A, eBioscience), IL-17 (TC11-18H10.1, Biolegend), IL-7Ra (SB/199, BD Biosciences), Ki-67 (MKI67, BD Biosciences), IL-33Ra (D1H9, Biolegend), Ly-6C (AL-21, BD Biosciences), Ly-6G (1A8, Biologend), MHC2 (M5/114.15.2, Biolegend), Pan-NK (DX-5, eBioscience), Podoplanin (8.1.1, Biolegend), Rorγt (AFKJS-9, eBioscience), TCR-β (H57-597, Biolegend), T-bet (4B10, Biolegend), Thy1.1 (OX-7, BD Biosciences), Thy1.2 (53-2.1, Biolegend), anti-human CD3 (UCHT1, Biolegend), CD4 (OKT4, Biolegend), CD8 (SK1, Biolegend), CD11b (M1/70, eBioscience), CD11c (3.9, Biolegend), CD19 (HIB19, Biolegend), CD45 (HI30, Biolegend), CD56 (HCD56, Biolegend), GM-CSF (BVD2-21C11, Biolegend), IFN-γ (4S.B3, Biolegend), IL-13 (JES10-5A2, Biolegend), IL-17 (BL168, Biolegend), PECy7-Streptavidin (BD Biosciences).

    Techniques: Staining, Mouse Assay, Injection, Adoptive Transfer Assay

    CSF-2 induces AM proliferation and rescues aberrant pulmonary surfactant PC accumulation in CD44 -/- mice. (A) Representative flow cytometry plots showing the proportion of BAL AMs from CD44 -/- mice after 7 days of PBS or CSF-2 treatment. (B) Number of AMs in the BAL from CD44 -/- mice after PBS or CSF-2 treatment. (C) BAL PC concentration from CD44 -/- mice after PBS or CSF-2 treatment. (D) Representative flow cytometry histograms comparing the phenotype of AMs from CD44 -/- mice after 7 days of PBS or CSF-2 treatment. (E) Graphs comparing the MFI of SSC-A, BODIPY, CD36, and CD11c between AMs from CD44 -/- mice after 7 days of PBS or CSF-2 treatment. Data show an average of two experiments ± SD, each with three to five mice. Background MFI of autofluorescence from unlabeled CD44 +/+ and CD44 -/- cells were respectively subtracted in the flow cytometry analysis. Significance indicated as * p

    Journal: bioRxiv

    Article Title: RNA sequencing and lipidomic analysis of alveolar macrophages from normal and CD44 deficient mice

    doi: 10.1101/845644

    Figure Lengend Snippet: CSF-2 induces AM proliferation and rescues aberrant pulmonary surfactant PC accumulation in CD44 -/- mice. (A) Representative flow cytometry plots showing the proportion of BAL AMs from CD44 -/- mice after 7 days of PBS or CSF-2 treatment. (B) Number of AMs in the BAL from CD44 -/- mice after PBS or CSF-2 treatment. (C) BAL PC concentration from CD44 -/- mice after PBS or CSF-2 treatment. (D) Representative flow cytometry histograms comparing the phenotype of AMs from CD44 -/- mice after 7 days of PBS or CSF-2 treatment. (E) Graphs comparing the MFI of SSC-A, BODIPY, CD36, and CD11c between AMs from CD44 -/- mice after 7 days of PBS or CSF-2 treatment. Data show an average of two experiments ± SD, each with three to five mice. Background MFI of autofluorescence from unlabeled CD44 +/+ and CD44 -/- cells were respectively subtracted in the flow cytometry analysis. Significance indicated as * p

    Article Snippet: Recombinant mouse CSF-2 (carrier free) was from BioLegend.

    Techniques: Mouse Assay, Flow Cytometry, Affinity Magnetic Separation, Concentration Assay

    PPARγ expression is defective in CD44 -/- AMs. (A) Representative confocal microscopy showing CD44 +/+ and CD44 -/- AMs labeled with intracellular PPARγ antibody and DAPI. (B) Comparison of nuclear PPARγ mean pixel intensity (MPI) between CD44 +/+ and CD44 -/- AMs, determined by confocal microscopy. (C) Representative flow cytometry histograms and graphs comparing intracellular PPARγ expression between CD44 +/+ and CD44 -/- AMs. (D) and (E) Representative flow cytometry histograms and graphs comparing the levels of CD36, BODIPY and CD11c levels by MFI (after subtraction of background autofluorescence) in CD44 +/+ AMs cultured with CSF-2 or CSF-2 and the PPARγ antagonist T0070907 (Antag) for 48 h in culture. Data show an average of three to five mice from each experiment ± SD, repeated twice or three times; for confocal imaging three to four fields containing cells were analyzed per mice. Significance indicated as * p

    Journal: bioRxiv

    Article Title: RNA sequencing and lipidomic analysis of alveolar macrophages from normal and CD44 deficient mice

    doi: 10.1101/845644

    Figure Lengend Snippet: PPARγ expression is defective in CD44 -/- AMs. (A) Representative confocal microscopy showing CD44 +/+ and CD44 -/- AMs labeled with intracellular PPARγ antibody and DAPI. (B) Comparison of nuclear PPARγ mean pixel intensity (MPI) between CD44 +/+ and CD44 -/- AMs, determined by confocal microscopy. (C) Representative flow cytometry histograms and graphs comparing intracellular PPARγ expression between CD44 +/+ and CD44 -/- AMs. (D) and (E) Representative flow cytometry histograms and graphs comparing the levels of CD36, BODIPY and CD11c levels by MFI (after subtraction of background autofluorescence) in CD44 +/+ AMs cultured with CSF-2 or CSF-2 and the PPARγ antagonist T0070907 (Antag) for 48 h in culture. Data show an average of three to five mice from each experiment ± SD, repeated twice or three times; for confocal imaging three to four fields containing cells were analyzed per mice. Significance indicated as * p

    Article Snippet: Recombinant mouse CSF-2 (carrier free) was from BioLegend.

    Techniques: Expressing, Affinity Magnetic Separation, Confocal Microscopy, Labeling, Flow Cytometry, Cell Culture, Mouse Assay, Imaging

    AMs lipid homeostasis is controlled by the extracellular milieu and the expression of CD44. (A) Schematic diagram showing donor AMs isolated from CD45.2 + CD44 -/- mice are transferred by i.t. into CD45.1 + CD44 +/+ host mice and analyzed 7 days later. (B) Representative flow cytometry plots comparing the SSC-A, BODIPY, CD36, and CD11c levels by CD45.2 + CD44 -/- donor AMs and CD45.1 + CD44 +/+ host AMs with control CD45.2 + CD44 -/- mice, 7 days after adoptive transfer. (C) Graphs comparing SSC-A, BODIPY, CD36, and CD11c MFI of CD45.2 + CD44 -/- donor AMs, CD45.1 + CD44 +/+ host AMs, and ex vivo CD44 -/- AMs. (D) Schematic diagram showing donor AMs isolated from CD45.1 + CD44 +/+ mice are transferred by i.t. into CD45.2 + CD44 -/- host mice and analyzed 7 days later. (E) Representative flow cytometry plots comparing the SSC-A, BODIPY, CD36, and CD11c levels in CD45.1 + CD44 +/+ donor AMs and CD45.2 + CD44 -/- host AMs compared with CD45.1 + CD44 +/+ control mice, 7 days after adoptive transfer. (F) Graphs comparing SSC-A, BODIPY, CD36, and CD11c MFI of CD45.1 + CD44 +/+ donor AMs, CD45.2 + CD44 -/- host AMs, and ex vivo CD44 +/+ AMs. (G) Representative flow cytometry histograms comparing the BODIPY and CD36 MFI of AMs after 48 h culture in CSF-2, or CSF-2 and PC. (H) and (I) Graphs comparing BODIPY labeling and CD36 expression by AMs cultured in CSF-2, or CSF-2 and PC for 48 h. For adoptive transfer experiments, data show an average of two experiments ± SD, each with three to five mice. Significance indicated as * p

    Journal: bioRxiv

    Article Title: RNA sequencing and lipidomic analysis of alveolar macrophages from normal and CD44 deficient mice

    doi: 10.1101/845644

    Figure Lengend Snippet: AMs lipid homeostasis is controlled by the extracellular milieu and the expression of CD44. (A) Schematic diagram showing donor AMs isolated from CD45.2 + CD44 -/- mice are transferred by i.t. into CD45.1 + CD44 +/+ host mice and analyzed 7 days later. (B) Representative flow cytometry plots comparing the SSC-A, BODIPY, CD36, and CD11c levels by CD45.2 + CD44 -/- donor AMs and CD45.1 + CD44 +/+ host AMs with control CD45.2 + CD44 -/- mice, 7 days after adoptive transfer. (C) Graphs comparing SSC-A, BODIPY, CD36, and CD11c MFI of CD45.2 + CD44 -/- donor AMs, CD45.1 + CD44 +/+ host AMs, and ex vivo CD44 -/- AMs. (D) Schematic diagram showing donor AMs isolated from CD45.1 + CD44 +/+ mice are transferred by i.t. into CD45.2 + CD44 -/- host mice and analyzed 7 days later. (E) Representative flow cytometry plots comparing the SSC-A, BODIPY, CD36, and CD11c levels in CD45.1 + CD44 +/+ donor AMs and CD45.2 + CD44 -/- host AMs compared with CD45.1 + CD44 +/+ control mice, 7 days after adoptive transfer. (F) Graphs comparing SSC-A, BODIPY, CD36, and CD11c MFI of CD45.1 + CD44 +/+ donor AMs, CD45.2 + CD44 -/- host AMs, and ex vivo CD44 +/+ AMs. (G) Representative flow cytometry histograms comparing the BODIPY and CD36 MFI of AMs after 48 h culture in CSF-2, or CSF-2 and PC. (H) and (I) Graphs comparing BODIPY labeling and CD36 expression by AMs cultured in CSF-2, or CSF-2 and PC for 48 h. For adoptive transfer experiments, data show an average of two experiments ± SD, each with three to five mice. Significance indicated as * p

    Article Snippet: Recombinant mouse CSF-2 (carrier free) was from BioLegend.

    Techniques: Affinity Magnetic Separation, Expressing, Isolation, Mouse Assay, Flow Cytometry, Adoptive Transfer Assay, Ex Vivo, Labeling, Cell Culture

    Role of TGF-β in the development of microglia in vitro. ( a and b ) MG400 analysis of (a) upregulated microglial genes and (b) downregulated microglial genes in cultured adult microglia in the presence of MCSF and TGF-β1 in comparison to adult microglia cultured in the presence of GM-CSF or MCSF alone (see Source data – Figure 6). One representative of three individual experiments is shown. (c) qPCR analysis of 6 selected microglial genes in cultured adult microglia in the presence of MCSF and TGF-β1. Gene expression level was normalized against Gapdh using ΔCt (n = 6, each biological triplicate consisted of two wells per treatment). Bars show mean normalized intensity ± s.e.m. * P

    Journal: Nature neuroscience

    Article Title: Identification of a Unique TGF-? Dependent Molecular and Functional Signature in Microglia

    doi: 10.1038/nn.3599

    Figure Lengend Snippet: Role of TGF-β in the development of microglia in vitro. ( a and b ) MG400 analysis of (a) upregulated microglial genes and (b) downregulated microglial genes in cultured adult microglia in the presence of MCSF and TGF-β1 in comparison to adult microglia cultured in the presence of GM-CSF or MCSF alone (see Source data – Figure 6). One representative of three individual experiments is shown. (c) qPCR analysis of 6 selected microglial genes in cultured adult microglia in the presence of MCSF and TGF-β1. Gene expression level was normalized against Gapdh using ΔCt (n = 6, each biological triplicate consisted of two wells per treatment). Bars show mean normalized intensity ± s.e.m. * P

    Article Snippet: Sorted microglia were cultured 6-well plate (1.2×105 cells/well in 3ml) or 12-well plate (6.5×104 cells/well in 2ml) in poly-D-lysine coated plates (BD Biosciences), and grown in microglia culture medium (DMEM/F-12 Glutamax; Invitrogen; supplemented with 10% FCS, 100U ml−1 penicillin, 100mg ml−1 streptomycin and mouse recombinant carrier free MCSF 10ng/ml (R & D Systems) or mouse recombinant carrier free GM-CSF 10ng ml−1 (R & D Systems) at 37°C, 5% CO2.

    Techniques: In Vitro, Cell Culture, Real-time Polymerase Chain Reaction, Expressing