Mouse GM-CSF R alpha APC-conjugated Antibody

Detection of GM‑CSF R alpha in J774A.1 Mouse Cell Line by Flow Cytometry.

J774A.1 mouse reticulum cell sarcoma macrophage cell line was stained with Rat Anti-Mouse GM-CSF Ra APC-conjugated Monoclonal Antibody (Catalog # FAB6130A, filled histogram) or isotype control antibody (Catalog # IC006A, open histogram). View our protocol for Staining Membrane-associated Proteins.

Detection of Mouse GM-CSFR alpha by Flow Cytometry

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, Ror gammat, 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 Il17aCre R26RFP635 SKG mice.(M) Quantitative RT-PCR analysis of Csf2 and Bhlhe40 in splenic naive CD25−CD44loCD4+ 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 × 106) 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 < 0.05, ∗∗p < 0.01. Data are representative of three independent experiments in (A)–(C), (E)–(J), (L), and (M) and pooled from more than two experiments in (D), (F), and (N). Vertical bars mean SD in (B), (E), (I), (K), (M), and (N). Image collected and cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S1074761318301468), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse GM-CSFR alpha by Flow Cytometry

GM-CSF from ILCs and Radio-Resistant Stromal Cells Is Crucial for Autoimmune Arthritis(A) Preparation of experimental groups for assessing arthritogenic effects of GM-CSF from ILCs or radio-resistant stromal cells. Rag2−/− or Csf2−/−Rag2−/− mice were x-irradiated (X-Rag2−/− mice) and transferred with BM cells from Csf2−/− or WT Rag2−/− mice. The resulting four groups of BM chimeras were transferred with CD4+ T cells from Csf2−/− SKG mice 6 weeks after BM reconstitution and assessed for arthritis development 12 weeks later.(B) Arthritis scores of four groups of mice shown in (A).(C) Proportion of total synovial ILCs from x-irradiated Rag2−/− mice reconstituted with Csf2−/− or WT Rag2−/− BM cells.(D) Flow cytometry of synovial ILCs for the expression of GM-CSF and IL-13 in arthritic joints of BM chimeras shown in (C).(E) Proportion of GM-CSF+IL-13−, GM-CSF+IL-13+, and GM-CSF−IL-13+ synovial ILCs (n = 6 each) as shown in (D).(F and G) The effects of ILC depletion on arthritis development. Thy1.1+Rag2−/− mice were x-irradiated and transferred with BM cells from Thy1.1+Rag2−/− and Thy1.2+Csf2−/− SKG mice. The resulting BM chimeras were i.p. injected with 20 mg mannan 6 weeks later, followed by i.v. injection with 500 μg anti-Thy1.1 mAb or control Rat IgG every week. Flow cytometry of synovial ILCs for GM-CSF and IL-13 expression (F). Arthritis scores in each group of mice monitored every week (G). Vertical bars mean SD (n = 8 each).∗p < 0.05, ∗∗p < 0.01. Data are representative of three independent experiments in (D) and (F), and pooled from three experiments in (B), (C), (E), and (G). Horizontal bars indicate the means in (B), (C), and (E). Image collected and cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S1074761318301468), licensed under a CC-BY license. Not internally tested by R&D Systems.