anti-f4 Search Results


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  • 93
    Thermo Fisher anti f4 80
    Anti F4 80, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 2930 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad rat anti mouse f4 80
    <t>F4/80-positive</t> cells from young animals enhance fracture repair and osteogenic differentiation. a Young (Y) and old (O) bone marrow cells were sorted into F4/80 high (+) and low (−) populations using flow cytometry and re-introduced into irradiated recipient old mice. Representative micro-CT radiographic images after a tibia fracture and Safranin O stained histologic images from the tibia of an old mouse 4 weeks after the bone was fractured in mice with transplanted with combinations of F4/80 high or low fractions from bone marrow from old or young donor mice. Graphs show bone volume/total volume (BV/TV) in %; total fibrous tissue/total volume (FV/TV) in %; bone plus calcified callus volume (mm 3 ) or total callous volume (mm 3 ), or relative density from tibias 4 weeks after fracture. Each data point is shown, along with means and 95% confidence intervals for each experimental group. n = 8 males, 9 females O−,O+, n = 9 males, 9 females for Y−,Y+, n = 9 males, 8 females for O−,Y+. b CFU-O from mice in A, with numbers that form from bone marrow cells from old mice. Representative cell culture plates (Von Kossa and Alk Phosphate staining shown) and each data point as well as means and 95% confidence intervals for each group with the mean from control culture defined as “1” (right). c Young (Y) and old (O) bone marrow cells sorted into F4/80 high (+) and low (−) populations using flow cytometry were re-introduced into irradiated recipient young mice ( n = 6 in each group). Representative radiographic images 2 and 4 weeks after the fracture, and histologic images from the tibia of old mice 4 weeks after the bone was fractured transplanted with various combinations of F4/80 high or low bone marrow fractions. Graphs show each data point for bone volume/total volume (BV/TV) in %; total fibrous tissue/total volume (FV/TV) in %; bone plus calcified callus volume (mm 3 ), total callous volume (mm 3 ), or relative density. Data are shown as means and 95% confidence intervals. n = 11 males, 10 females for Y−;O+, n = 10 males, 10 females for O−:Y+, n = 10 males, 11 females for Y−;Y+. An asterisk over a data point shows a significance p
    Rat Anti Mouse F4 80, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1945 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti f4 80
    Depletion of Macrophages Reduces Tendon Adhesion In Vivo (A) Experimental scheme showing the application of Clo- or PBS-containing liposomes in mice with TI or SO. (B) Macroscopic images of FDL tendons and adhesion grading score (n = 5 per group). (C) Maximal tensile strength and MTP joint flexion ROM (n = 5 per group). (D) Immunostaining of macrophages <t>(F4/80</t> + cells, green) in each group; cell nuclei were stained with DAPI (blue). T, tendon; M, muscle. White arrows indicate F4/80-positive cells. Dotted lines indicate the border of FDL tendon or surrounding tissue. Scale bars, 50 μm. (E and F) Histology of adhesion. T, tendon; B, bone; M, muscle. Arrows indicate tendon adhesion in (E) H E and (F) Masson staining. Dotted lines indicate the border of FDL tendon or surrounding tissue. Scale bars, 50 μm. (G) Histological adhesion score and histological healing score (n = 5 per group). (H) Western blot analysis and quantification of the levels of COL I, COL III, α-SMA, and TGF-β1 expression (n = 3 per group). *p
    Anti F4 80, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1025 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher f4 80 monoclonal antibody
    Depletion of Macrophages Reduces Tendon Adhesion In Vivo (A) Experimental scheme showing the application of Clo- or PBS-containing liposomes in mice with TI or SO. (B) Macroscopic images of FDL tendons and adhesion grading score (n = 5 per group). (C) Maximal tensile strength and MTP joint flexion ROM (n = 5 per group). (D) Immunostaining of macrophages <t>(F4/80</t> + cells, green) in each group; cell nuclei were stained with DAPI (blue). T, tendon; M, muscle. White arrows indicate F4/80-positive cells. Dotted lines indicate the border of FDL tendon or surrounding tissue. Scale bars, 50 μm. (E and F) Histology of adhesion. T, tendon; B, bone; M, muscle. Arrows indicate tendon adhesion in (E) H E and (F) Masson staining. Dotted lines indicate the border of FDL tendon or surrounding tissue. Scale bars, 50 μm. (G) Histological adhesion score and histological healing score (n = 5 per group). (H) Western blot analysis and quantification of the levels of COL I, COL III, α-SMA, and TGF-β1 expression (n = 3 per group). *p
    F4 80 Monoclonal Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1982 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam rat anti f4 80
    Pentoxifylline treatment reduced inflammation. (A) Immunohistochemistry for <t>F4/80</t> was performed on mice peritoneal tissue in the different groups at day 3 and day 7. The F4/80 expression was increased in the PA group. Representative images of the sham group, the PA group, and the PA+PTX group are shown (Original magnification, x200, bar = 100 µm). (B) Quantification of F4/80 + cells (%) in high-powered field (HPF) at x400 magnification. Data are expressed as the mean± SE. ∗ P
    Rat Anti F4 80, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 610 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    f4 80  (Abcam)
    99
    Abcam f4 80
    Pentoxifylline treatment reduced inflammation. (A) Immunohistochemistry for <t>F4/80</t> was performed on mice peritoneal tissue in the different groups at day 3 and day 7. The F4/80 expression was increased in the PA group. Representative images of the sham group, the PA group, and the PA+PTX group are shown (Original magnification, x200, bar = 100 µm). (B) Quantification of F4/80 + cells (%) in high-powered field (HPF) at x400 magnification. Data are expressed as the mean± SE. ∗ P
    F4 80, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 3492 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rat anti f4 80
    Lack of substantial inflammatory cell infiltration into lung in mice following intravenous injection of GAd. Immunofluorescence microscopy was used to analyze <t>F4/80</t> + macrophages and CD8 + T cells in lungs from untreated C57BL/6J mice (untreated, n = 3) and from mice at days 1, 3, 7, and 11 following intravenous injection of 1.0 × 10 11 viral particles (vp) of HAd5 and GAd ( n = 2 for each virus and time point). Magnifications, ×100. Red, CD31/endomucin; green, GFP; blue, DAPI.
    Rat Anti F4 80, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 270 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend anti f4 80
    Lack of substantial inflammatory cell infiltration into lung in mice following intravenous injection of GAd. Immunofluorescence microscopy was used to analyze <t>F4/80</t> + macrophages and CD8 + T cells in lungs from untreated C57BL/6J mice (untreated, n = 3) and from mice at days 1, 3, 7, and 11 following intravenous injection of 1.0 × 10 11 viral particles (vp) of HAd5 and GAd ( n = 2 for each virus and time point). Magnifications, ×100. Red, CD31/endomucin; green, GFP; blue, DAPI.
    Anti F4 80, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 1473 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend pe anti f4 80
    Exposure to FA21 MWCNT exacerbates HDM-induced allergic lung inflammation and promotes cysLT production. To examine the effect of MWCNT exposure on pre-existing allergic airway inflammation, C57BL/6 mice (4–6 per group) were intranasally challenged with PBS (control) or HDM allergen over a 2-week period (on days 0, 7, and 14) to induce allergic inflammation and then exposed to FA21 MWCNT (50 μg) on day 15 by intratracheal administration. BALF and lung tissue were collected 24 h after MWCNT administration on day 16 for analysis of pulmonary inflammation (as illustrated in the experimental design in Figure 1 ). Groups comprised of mice treated with FA21, HDM, or FA21 + HDM. Control mice were treated with carrier alone. (A) BALF cell differential counts were determined and expressed as absolute cell number per mouse of lymphocytes (LYM), eosinophils (EOS), macrophages (MAC), and polymorphonuclear neutrophils (NEU). (B) Cell-associated EPO levels in the BALF were determined by colorimetric assay. (C) CD11b + <t>F4/80</t> - GR1 - Siglec-F + eosinophil numbers present in the BALF were analyzed by flow cytometry. (D) Peribronchial and perivascular inflammation was assessed by histological analysis of lung tissue sections stained with H E (20×). In addition, inflammatory cells infiltrating the airways were examined by microscopic evaluation and imaging of cytospin preparations of BALF cells stained using Hema3. (E) CysLT levels present in BALF determined by ELISA. Results are mean ± SEM ( n = 6), ∗∗ p
    Pe Anti F4 80, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson anti f4 80
    CRACC-CRACC interaction between macrophages and NK cells enhances the cytokine secretion in vitro. A , Ana-1 cells pre-transfected with liposome encapsulated siNeg or siCRACC was subsequently stimulated with Poly I:C. The expression of CRACC mRNA was assayed by quantitative PCR at 3h time point post the Poly I:C stimulation. B , C D , Ana-1 cells pre-transfected with nanoparticle encapsulated siNeg or siCRACC was cultured in absence or presence of NK cells, which were freshly isolated from mice pre-treated by Poly I:C. Then Ana-1 cells cultured with or without NK cells were stimulated with Poly I:C; and D-GalN was made as control. CRACC expression on Ana-1 cells was analyzed by flow cytometry ( B ) and the cytokine (TNF-α and IFN-γ) in the culture supernatant was assayed by ELISA ( C , D ) at 24h time point post the Poly I:C or D-GalN stimulation; Ana-1 cells were identified as <t>F4/80</t> + cells. E , Ana-1 cells were cocultured with freshly isolated NK cells from mice pre-treated by Poly I:C; and Ana-1 cells alone or NK cells alone were made as control. The Ana-1-NK coculture was stimulated by Poly I:C in the presence of isotype control Ab (Rat IgG), CRACC mAb (anti-CRACC), or NKG2D mAb (anti-NKG2D), respectively. The IFN-γ in culture supernatant was assayed by ELISA at 24h time point post the Poly I:C stimulation. *P
    Anti F4 80, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 691 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend apc anti f4 80
    CRACC-CRACC interaction between macrophages and NK cells enhances the cytokine secretion in vitro. A , Ana-1 cells pre-transfected with liposome encapsulated siNeg or siCRACC was subsequently stimulated with Poly I:C. The expression of CRACC mRNA was assayed by quantitative PCR at 3h time point post the Poly I:C stimulation. B , C D , Ana-1 cells pre-transfected with nanoparticle encapsulated siNeg or siCRACC was cultured in absence or presence of NK cells, which were freshly isolated from mice pre-treated by Poly I:C. Then Ana-1 cells cultured with or without NK cells were stimulated with Poly I:C; and D-GalN was made as control. CRACC expression on Ana-1 cells was analyzed by flow cytometry ( B ) and the cytokine (TNF-α and IFN-γ) in the culture supernatant was assayed by ELISA ( C , D ) at 24h time point post the Poly I:C or D-GalN stimulation; Ana-1 cells were identified as <t>F4/80</t> + cells. E , Ana-1 cells were cocultured with freshly isolated NK cells from mice pre-treated by Poly I:C; and Ana-1 cells alone or NK cells alone were made as control. The Ana-1-NK coculture was stimulated by Poly I:C in the presence of isotype control Ab (Rat IgG), CRACC mAb (anti-CRACC), or NKG2D mAb (anti-NKG2D), respectively. The IFN-γ in culture supernatant was assayed by ELISA at 24h time point post the Poly I:C stimulation. *P
    Apc Anti F4 80, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 68 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti f4 80 antibody ci a3 1
    CRACC-CRACC interaction between macrophages and NK cells enhances the cytokine secretion in vitro. A , Ana-1 cells pre-transfected with liposome encapsulated siNeg or siCRACC was subsequently stimulated with Poly I:C. The expression of CRACC mRNA was assayed by quantitative PCR at 3h time point post the Poly I:C stimulation. B , C D , Ana-1 cells pre-transfected with nanoparticle encapsulated siNeg or siCRACC was cultured in absence or presence of NK cells, which were freshly isolated from mice pre-treated by Poly I:C. Then Ana-1 cells cultured with or without NK cells were stimulated with Poly I:C; and D-GalN was made as control. CRACC expression on Ana-1 cells was analyzed by flow cytometry ( B ) and the cytokine (TNF-α and IFN-γ) in the culture supernatant was assayed by ELISA ( C , D ) at 24h time point post the Poly I:C or D-GalN stimulation; Ana-1 cells were identified as <t>F4/80</t> + cells. E , Ana-1 cells were cocultured with freshly isolated NK cells from mice pre-treated by Poly I:C; and Ana-1 cells alone or NK cells alone were made as control. The Ana-1-NK coculture was stimulated by Poly I:C in the presence of isotype control Ab (Rat IgG), CRACC mAb (anti-CRACC), or NKG2D mAb (anti-NKG2D), respectively. The IFN-γ in culture supernatant was assayed by ELISA at 24h time point post the Poly I:C stimulation. *P
    Anti F4 80 Antibody Ci A3 1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 125 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend fitc anti f4 80
    CRACC-CRACC interaction between macrophages and NK cells enhances the cytokine secretion in vitro. A , Ana-1 cells pre-transfected with liposome encapsulated siNeg or siCRACC was subsequently stimulated with Poly I:C. The expression of CRACC mRNA was assayed by quantitative PCR at 3h time point post the Poly I:C stimulation. B , C D , Ana-1 cells pre-transfected with nanoparticle encapsulated siNeg or siCRACC was cultured in absence or presence of NK cells, which were freshly isolated from mice pre-treated by Poly I:C. Then Ana-1 cells cultured with or without NK cells were stimulated with Poly I:C; and D-GalN was made as control. CRACC expression on Ana-1 cells was analyzed by flow cytometry ( B ) and the cytokine (TNF-α and IFN-γ) in the culture supernatant was assayed by ELISA ( C , D ) at 24h time point post the Poly I:C or D-GalN stimulation; Ana-1 cells were identified as <t>F4/80</t> + cells. E , Ana-1 cells were cocultured with freshly isolated NK cells from mice pre-treated by Poly I:C; and Ana-1 cells alone or NK cells alone were made as control. The Ana-1-NK coculture was stimulated by Poly I:C in the presence of isotype control Ab (Rat IgG), CRACC mAb (anti-CRACC), or NKG2D mAb (anti-NKG2D), respectively. The IFN-γ in culture supernatant was assayed by ELISA at 24h time point post the Poly I:C stimulation. *P
    Fitc Anti F4 80, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher anti f4 80 apc
    CRACC-CRACC interaction between macrophages and NK cells enhances the cytokine secretion in vitro. A , Ana-1 cells pre-transfected with liposome encapsulated siNeg or siCRACC was subsequently stimulated with Poly I:C. The expression of CRACC mRNA was assayed by quantitative PCR at 3h time point post the Poly I:C stimulation. B , C D , Ana-1 cells pre-transfected with nanoparticle encapsulated siNeg or siCRACC was cultured in absence or presence of NK cells, which were freshly isolated from mice pre-treated by Poly I:C. Then Ana-1 cells cultured with or without NK cells were stimulated with Poly I:C; and D-GalN was made as control. CRACC expression on Ana-1 cells was analyzed by flow cytometry ( B ) and the cytokine (TNF-α and IFN-γ) in the culture supernatant was assayed by ELISA ( C , D ) at 24h time point post the Poly I:C or D-GalN stimulation; Ana-1 cells were identified as <t>F4/80</t> + cells. E , Ana-1 cells were cocultured with freshly isolated NK cells from mice pre-treated by Poly I:C; and Ana-1 cells alone or NK cells alone were made as control. The Ana-1-NK coculture was stimulated by Poly I:C in the presence of isotype control Ab (Rat IgG), CRACC mAb (anti-CRACC), or NKG2D mAb (anti-NKG2D), respectively. The IFN-γ in culture supernatant was assayed by ELISA at 24h time point post the Poly I:C stimulation. *P
    Anti F4 80 Apc, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 585 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Santa Cruz Biotechnology anti f4 80
    CLEC4F + cells were appeared in the liver environment under Kupffer cell depletion and inflammatory stage. (A) Kupffer cells were depleted by Cl 2 MBP-encapsulated liposome by intravenous injection (100 µl/mouse) at day 0 and livers were harvest at day 1, 7, 14 and 28. <t>F4/80</t> and CLEC4F immunohistochemistry of liver sections were performed. (B) The numbers of F4/80 + or CLEC4F + cells in livers were shown. For generating inflammatory stage, wild-type and Clec4f−/− littermates were infected with L. monocytogenes (1×10 5 CFU/mouse) intravenously. (C) The numbers of F4/80 + or CLEC4F + cells in livers during L. monocytogenes infection. (D) Immunohistochemistry of L. monocytogenes infected livers of wild-type and Clec4f−/− mice at day 5 after infection. (E) Kaplan-Meier survival curves were shown for Clec4f−/− or wild-type littermates with L. monocytogenes infection. The p value was determined by Log-rank test.
    Anti F4 80, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 476 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    F4/80-positive cells from young animals enhance fracture repair and osteogenic differentiation. a Young (Y) and old (O) bone marrow cells were sorted into F4/80 high (+) and low (−) populations using flow cytometry and re-introduced into irradiated recipient old mice. Representative micro-CT radiographic images after a tibia fracture and Safranin O stained histologic images from the tibia of an old mouse 4 weeks after the bone was fractured in mice with transplanted with combinations of F4/80 high or low fractions from bone marrow from old or young donor mice. Graphs show bone volume/total volume (BV/TV) in %; total fibrous tissue/total volume (FV/TV) in %; bone plus calcified callus volume (mm 3 ) or total callous volume (mm 3 ), or relative density from tibias 4 weeks after fracture. Each data point is shown, along with means and 95% confidence intervals for each experimental group. n = 8 males, 9 females O−,O+, n = 9 males, 9 females for Y−,Y+, n = 9 males, 8 females for O−,Y+. b CFU-O from mice in A, with numbers that form from bone marrow cells from old mice. Representative cell culture plates (Von Kossa and Alk Phosphate staining shown) and each data point as well as means and 95% confidence intervals for each group with the mean from control culture defined as “1” (right). c Young (Y) and old (O) bone marrow cells sorted into F4/80 high (+) and low (−) populations using flow cytometry were re-introduced into irradiated recipient young mice ( n = 6 in each group). Representative radiographic images 2 and 4 weeks after the fracture, and histologic images from the tibia of old mice 4 weeks after the bone was fractured transplanted with various combinations of F4/80 high or low bone marrow fractions. Graphs show each data point for bone volume/total volume (BV/TV) in %; total fibrous tissue/total volume (FV/TV) in %; bone plus calcified callus volume (mm 3 ), total callous volume (mm 3 ), or relative density. Data are shown as means and 95% confidence intervals. n = 11 males, 10 females for Y−;O+, n = 10 males, 10 females for O−:Y+, n = 10 males, 11 females for Y−;Y+. An asterisk over a data point shows a significance p

    Journal: Nature Communications

    Article Title: Macrophage cells secrete factors including LRP1 that orchestrate the rejuvenation of bone repair in mice

    doi: 10.1038/s41467-018-07666-0

    Figure Lengend Snippet: F4/80-positive cells from young animals enhance fracture repair and osteogenic differentiation. a Young (Y) and old (O) bone marrow cells were sorted into F4/80 high (+) and low (−) populations using flow cytometry and re-introduced into irradiated recipient old mice. Representative micro-CT radiographic images after a tibia fracture and Safranin O stained histologic images from the tibia of an old mouse 4 weeks after the bone was fractured in mice with transplanted with combinations of F4/80 high or low fractions from bone marrow from old or young donor mice. Graphs show bone volume/total volume (BV/TV) in %; total fibrous tissue/total volume (FV/TV) in %; bone plus calcified callus volume (mm 3 ) or total callous volume (mm 3 ), or relative density from tibias 4 weeks after fracture. Each data point is shown, along with means and 95% confidence intervals for each experimental group. n = 8 males, 9 females O−,O+, n = 9 males, 9 females for Y−,Y+, n = 9 males, 8 females for O−,Y+. b CFU-O from mice in A, with numbers that form from bone marrow cells from old mice. Representative cell culture plates (Von Kossa and Alk Phosphate staining shown) and each data point as well as means and 95% confidence intervals for each group with the mean from control culture defined as “1” (right). c Young (Y) and old (O) bone marrow cells sorted into F4/80 high (+) and low (−) populations using flow cytometry were re-introduced into irradiated recipient young mice ( n = 6 in each group). Representative radiographic images 2 and 4 weeks after the fracture, and histologic images from the tibia of old mice 4 weeks after the bone was fractured transplanted with various combinations of F4/80 high or low bone marrow fractions. Graphs show each data point for bone volume/total volume (BV/TV) in %; total fibrous tissue/total volume (FV/TV) in %; bone plus calcified callus volume (mm 3 ), total callous volume (mm 3 ), or relative density. Data are shown as means and 95% confidence intervals. n = 11 males, 10 females for Y−;O+, n = 10 males, 10 females for O−:Y+, n = 10 males, 11 females for Y−;Y+. An asterisk over a data point shows a significance p

    Article Snippet: Immunohistochemistry and histomorphometry For F4/80 immunohistochemistry, antigen retrieval was performed with 20 μg/ml of proteinase K for 10 min and stained with primary rat- anti-mouse F4/80 (AbD Serotec, catalogue number MCA497) antibody diluted 1:100.

    Techniques: Flow Cytometry, Cytometry, Irradiation, Mouse Assay, Micro-CT, Staining, Cell Culture

    Immunohistochemistry for F4/80 (total macrophages ( a ), iNOS (M1 macrophages ( b ) and arginase-1 (M2 macrophages ( c ). C: control; ELA: elastase-induced emphysema; Sal: i.p. injection of saline; Ghr: i.p. injection of ghrelin. Boxes show the interquartile range (25 th –75 th percentile), whiskers encompass the range (minimum–maximum), and horizontal lines represent the median in 10 animals/group. * vs . C-Sal. # vs . ELA-Sal

    Journal: Respiratory Research

    Article Title: Ghrelin therapy improves lung and cardiovascular function in experimental emphysema

    doi: 10.1186/s12931-017-0668-9

    Figure Lengend Snippet: Immunohistochemistry for F4/80 (total macrophages ( a ), iNOS (M1 macrophages ( b ) and arginase-1 (M2 macrophages ( c ). C: control; ELA: elastase-induced emphysema; Sal: i.p. injection of saline; Ghr: i.p. injection of ghrelin. Boxes show the interquartile range (25 th –75 th percentile), whiskers encompass the range (minimum–maximum), and horizontal lines represent the median in 10 animals/group. * vs . C-Sal. # vs . ELA-Sal

    Article Snippet: Immunohistochemistry Immunohistochemical analysis for alveolar macrophages was characterized using F4/80 antibody (1:50; MCA497, AbD Serotec, Raleigh, NC, USA).

    Techniques: Immunohistochemistry, Injection

    Depletion of Macrophages Reduces Tendon Adhesion In Vivo (A) Experimental scheme showing the application of Clo- or PBS-containing liposomes in mice with TI or SO. (B) Macroscopic images of FDL tendons and adhesion grading score (n = 5 per group). (C) Maximal tensile strength and MTP joint flexion ROM (n = 5 per group). (D) Immunostaining of macrophages (F4/80 + cells, green) in each group; cell nuclei were stained with DAPI (blue). T, tendon; M, muscle. White arrows indicate F4/80-positive cells. Dotted lines indicate the border of FDL tendon or surrounding tissue. Scale bars, 50 μm. (E and F) Histology of adhesion. T, tendon; B, bone; M, muscle. Arrows indicate tendon adhesion in (E) H E and (F) Masson staining. Dotted lines indicate the border of FDL tendon or surrounding tissue. Scale bars, 50 μm. (G) Histological adhesion score and histological healing score (n = 5 per group). (H) Western blot analysis and quantification of the levels of COL I, COL III, α-SMA, and TGF-β1 expression (n = 3 per group). *p

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Macrophage-Derived miRNA-Containing Exosomes Induce Peritendinous Fibrosis after Tendon Injury through the miR-21-5p/Smad7 Pathway

    doi: 10.1016/j.omtn.2018.11.006

    Figure Lengend Snippet: Depletion of Macrophages Reduces Tendon Adhesion In Vivo (A) Experimental scheme showing the application of Clo- or PBS-containing liposomes in mice with TI or SO. (B) Macroscopic images of FDL tendons and adhesion grading score (n = 5 per group). (C) Maximal tensile strength and MTP joint flexion ROM (n = 5 per group). (D) Immunostaining of macrophages (F4/80 + cells, green) in each group; cell nuclei were stained with DAPI (blue). T, tendon; M, muscle. White arrows indicate F4/80-positive cells. Dotted lines indicate the border of FDL tendon or surrounding tissue. Scale bars, 50 μm. (E and F) Histology of adhesion. T, tendon; B, bone; M, muscle. Arrows indicate tendon adhesion in (E) H E and (F) Masson staining. Dotted lines indicate the border of FDL tendon or surrounding tissue. Scale bars, 50 μm. (G) Histological adhesion score and histological healing score (n = 5 per group). (H) Western blot analysis and quantification of the levels of COL I, COL III, α-SMA, and TGF-β1 expression (n = 3 per group). *p

    Article Snippet: The primary antibody used in this study was anti-F4/80 (1:200, Abcam, ab6640).

    Techniques: In Vivo, Mouse Assay, Immunostaining, Staining, Western Blot, Expressing

    CNF1 reduces CD36 expression in RAW264.7. (A) qRT-PCR analysis of CD36 mRNA and protein levels in RAW264.7 treated with CNF1 (3 nM), C866S (3 nM), and PBS for 3, 6, and 12 h. (B) FACS analysis of CD36 expression in RAW264.7 treated with CNF1 (3 nM), C866S (3 nM), PBS and LPS (1 μg/ml, positive control) for 12 h. (C) Western blotting analysis of CD36 protein levels in RAW264.7 treated with CNF1 (3 nM), C866S (3 nM), and PBS for 3, 6, and 12 h. (D) Immunofluorescence analysis of CD36 expression in RAW264.7 treated with CNF1 (3 nM), C866S (3 nM), PBS, and LPS (1 μg/ml, positive control) for 6 h. Scale bar, 20 μm. (E) Immunofluorescence analysis of CD36 expression in macrophages of bladder tissues infected by CNF1-expressing CFT073 and vector control CFT073 at 24 hpi, and by UTI89 and Δcnf1 at 48 hpi, respectively. Scale bar, 20 μm. Blue, nucleus; red, F4/80; Green, CD36. Data are from three combined independent experiments each with two replicate wells ( n = 6) (A) . Data are from four combined independent experiments (B) . Quantitative analysis of CD36 signal is using Image-Pro Plus, and data are from three combined independent experiments each with two fields ( n = 6) (D,E) . Data are the mean ± SD, One-way ANOVA, * P

    Journal: Frontiers in Immunology

    Article Title: Cytotoxic Necrotizing Factor 1 Downregulates CD36 Transcription in Macrophages to Induce Inflammation During Acute Urinary Tract Infections

    doi: 10.3389/fimmu.2018.01987

    Figure Lengend Snippet: CNF1 reduces CD36 expression in RAW264.7. (A) qRT-PCR analysis of CD36 mRNA and protein levels in RAW264.7 treated with CNF1 (3 nM), C866S (3 nM), and PBS for 3, 6, and 12 h. (B) FACS analysis of CD36 expression in RAW264.7 treated with CNF1 (3 nM), C866S (3 nM), PBS and LPS (1 μg/ml, positive control) for 12 h. (C) Western blotting analysis of CD36 protein levels in RAW264.7 treated with CNF1 (3 nM), C866S (3 nM), and PBS for 3, 6, and 12 h. (D) Immunofluorescence analysis of CD36 expression in RAW264.7 treated with CNF1 (3 nM), C866S (3 nM), PBS, and LPS (1 μg/ml, positive control) for 6 h. Scale bar, 20 μm. (E) Immunofluorescence analysis of CD36 expression in macrophages of bladder tissues infected by CNF1-expressing CFT073 and vector control CFT073 at 24 hpi, and by UTI89 and Δcnf1 at 48 hpi, respectively. Scale bar, 20 μm. Blue, nucleus; red, F4/80; Green, CD36. Data are from three combined independent experiments each with two replicate wells ( n = 6) (A) . Data are from four combined independent experiments (B) . Quantitative analysis of CD36 signal is using Image-Pro Plus, and data are from three combined independent experiments each with two fields ( n = 6) (D,E) . Data are the mean ± SD, One-way ANOVA, * P

    Article Snippet: Frozen sections (5 μm) were cut and air-dried at room temperature for 20 min and fixed with cold acetone for 10 min. Then the tissues were immediately submerged into methanol for 20 min and 3% hydrogen peroxide in methanol for 10 min. After rehydration in PBS, sections were blocked with 5% BSA for 1 h, incubated with F4/80 antibody (Abcam, ab6640, 1:100), CD36 antibody (Proteintech, 18836-1-AP, 1:100) in blocking buffer overnight at 4°C.

    Techniques: Expressing, Quantitative RT-PCR, FACS, Positive Control, Western Blot, Immunofluorescence, Infection, Plasmid Preparation

    Pentoxifylline treatment reduced inflammation. (A) Immunohistochemistry for F4/80 was performed on mice peritoneal tissue in the different groups at day 3 and day 7. The F4/80 expression was increased in the PA group. Representative images of the sham group, the PA group, and the PA+PTX group are shown (Original magnification, x200, bar = 100 µm). (B) Quantification of F4/80 + cells (%) in high-powered field (HPF) at x400 magnification. Data are expressed as the mean± SE. ∗ P

    Journal: PeerJ

    Article Title: Pentoxifylline decreases post-operative intra-abdominal adhesion formation in an animal model

    doi: 10.7717/peerj.5434

    Figure Lengend Snippet: Pentoxifylline treatment reduced inflammation. (A) Immunohistochemistry for F4/80 was performed on mice peritoneal tissue in the different groups at day 3 and day 7. The F4/80 expression was increased in the PA group. Representative images of the sham group, the PA group, and the PA+PTX group are shown (Original magnification, x200, bar = 100 µm). (B) Quantification of F4/80 + cells (%) in high-powered field (HPF) at x400 magnification. Data are expressed as the mean± SE. ∗ P

    Article Snippet: Sections were incubated with primary antibodies, rabbit anti-ki67 (1:200, Abcam, Cambridge, MA, USA), rat anti-F4/80 (1:200, Abcam) or rabbit anti-FSP-1 (1:200, Abcam) overnight at 4 °C.

    Techniques: Immunohistochemistry, Mouse Assay, Expressing

    Lack of substantial inflammatory cell infiltration into lung in mice following intravenous injection of GAd. Immunofluorescence microscopy was used to analyze F4/80 + macrophages and CD8 + T cells in lungs from untreated C57BL/6J mice (untreated, n = 3) and from mice at days 1, 3, 7, and 11 following intravenous injection of 1.0 × 10 11 viral particles (vp) of HAd5 and GAd ( n = 2 for each virus and time point). Magnifications, ×100. Red, CD31/endomucin; green, GFP; blue, DAPI.

    Journal: Journal of Virology

    Article Title: A New Gorilla Adenoviral Vector with Natural Lung Tropism Avoids Liver Toxicity and Is Amenable to Capsid Engineering and Vector Retargeting

    doi: 10.1128/JVI.00265-20

    Figure Lengend Snippet: Lack of substantial inflammatory cell infiltration into lung in mice following intravenous injection of GAd. Immunofluorescence microscopy was used to analyze F4/80 + macrophages and CD8 + T cells in lungs from untreated C57BL/6J mice (untreated, n = 3) and from mice at days 1, 3, 7, and 11 following intravenous injection of 1.0 × 10 11 viral particles (vp) of HAd5 and GAd ( n = 2 for each virus and time point). Magnifications, ×100. Red, CD31/endomucin; green, GFP; blue, DAPI.

    Article Snippet: The primary antibodies used in this study included rat anti-endomucin (1:1,000; catalog number 14-5851-81; eBioscience), Armenian hamster anti-CD31 (1:1,000; catalog number MAB1398Z; MilliporeSigma), chicken anti-GFP (1:400; catalog number A10262; Thermo Fisher Scientific), rabbit anti-mCherry (1:250; catalog number ab213511; Abcam), hamster anti-podoplanin (1:100; catalog number 8.1.1; Developmental Studies Hybridoma Bank), rabbit anti-ProSP-C (1:1,000; catalog number AB3786; MilliporeSigma), rabbit anti-α-SMA (1:100; catalog number ab5694; Abcam), goat antivimentin (1:20; catalog number sc7557; Santa Cruz Biotechnology), rat anti-CD45 (1:25; catalog number 550539; BD Biosciences), rat anti-F4/80 (1:50; catalog number MA1-91124; Thermo Fisher Scientific), rat anti-CD8 (1:20; catalog number 550281; BD Biosciences), and rabbit anti-activated caspase 3 (1:400; catalog number 9661; Cell Signaling Technology).

    Techniques: Mouse Assay, Injection, Immunofluorescence, Microscopy

    Rapid kinetics of liver Kupffer cell depletion followed by low levels of inflammatory cell infiltration to liver in mice following intravenous injection of GAd. Immunofluorescence microscopy analysis of F4/80 + macrophages (top two rows) and CD8 + T cells (bottom two rows) in liver from untreated C57BL/6J mice (untreated, n = 3) and from mice at days 1, 3, 5, 7, and 11 following intravenous injection of 1.0 × 10 11 viral particles (vp) of HAd5 and GAd ( n = 2 for each virus and time point). Magnifications, ×100. Red, CD31/endomucin; green, GFP; blue, DAPI.

    Journal: Journal of Virology

    Article Title: A New Gorilla Adenoviral Vector with Natural Lung Tropism Avoids Liver Toxicity and Is Amenable to Capsid Engineering and Vector Retargeting

    doi: 10.1128/JVI.00265-20

    Figure Lengend Snippet: Rapid kinetics of liver Kupffer cell depletion followed by low levels of inflammatory cell infiltration to liver in mice following intravenous injection of GAd. Immunofluorescence microscopy analysis of F4/80 + macrophages (top two rows) and CD8 + T cells (bottom two rows) in liver from untreated C57BL/6J mice (untreated, n = 3) and from mice at days 1, 3, 5, 7, and 11 following intravenous injection of 1.0 × 10 11 viral particles (vp) of HAd5 and GAd ( n = 2 for each virus and time point). Magnifications, ×100. Red, CD31/endomucin; green, GFP; blue, DAPI.

    Article Snippet: The primary antibodies used in this study included rat anti-endomucin (1:1,000; catalog number 14-5851-81; eBioscience), Armenian hamster anti-CD31 (1:1,000; catalog number MAB1398Z; MilliporeSigma), chicken anti-GFP (1:400; catalog number A10262; Thermo Fisher Scientific), rabbit anti-mCherry (1:250; catalog number ab213511; Abcam), hamster anti-podoplanin (1:100; catalog number 8.1.1; Developmental Studies Hybridoma Bank), rabbit anti-ProSP-C (1:1,000; catalog number AB3786; MilliporeSigma), rabbit anti-α-SMA (1:100; catalog number ab5694; Abcam), goat antivimentin (1:20; catalog number sc7557; Santa Cruz Biotechnology), rat anti-CD45 (1:25; catalog number 550539; BD Biosciences), rat anti-F4/80 (1:50; catalog number MA1-91124; Thermo Fisher Scientific), rat anti-CD8 (1:20; catalog number 550281; BD Biosciences), and rabbit anti-activated caspase 3 (1:400; catalog number 9661; Cell Signaling Technology).

    Techniques: Mouse Assay, Injection, Immunofluorescence, Microscopy

    Exposure to FA21 MWCNT exacerbates HDM-induced allergic lung inflammation and promotes cysLT production. To examine the effect of MWCNT exposure on pre-existing allergic airway inflammation, C57BL/6 mice (4–6 per group) were intranasally challenged with PBS (control) or HDM allergen over a 2-week period (on days 0, 7, and 14) to induce allergic inflammation and then exposed to FA21 MWCNT (50 μg) on day 15 by intratracheal administration. BALF and lung tissue were collected 24 h after MWCNT administration on day 16 for analysis of pulmonary inflammation (as illustrated in the experimental design in Figure 1 ). Groups comprised of mice treated with FA21, HDM, or FA21 + HDM. Control mice were treated with carrier alone. (A) BALF cell differential counts were determined and expressed as absolute cell number per mouse of lymphocytes (LYM), eosinophils (EOS), macrophages (MAC), and polymorphonuclear neutrophils (NEU). (B) Cell-associated EPO levels in the BALF were determined by colorimetric assay. (C) CD11b + F4/80 - GR1 - Siglec-F + eosinophil numbers present in the BALF were analyzed by flow cytometry. (D) Peribronchial and perivascular inflammation was assessed by histological analysis of lung tissue sections stained with H E (20×). In addition, inflammatory cells infiltrating the airways were examined by microscopic evaluation and imaging of cytospin preparations of BALF cells stained using Hema3. (E) CysLT levels present in BALF determined by ELISA. Results are mean ± SEM ( n = 6), ∗∗ p

    Journal: Frontiers in Pharmacology

    Article Title: Multi-Walled Carbon Nanotubes Augment Allergic Airway Eosinophilic Inflammation by Promoting Cysteinyl Leukotriene Production

    doi: 10.3389/fphar.2018.00585

    Figure Lengend Snippet: Exposure to FA21 MWCNT exacerbates HDM-induced allergic lung inflammation and promotes cysLT production. To examine the effect of MWCNT exposure on pre-existing allergic airway inflammation, C57BL/6 mice (4–6 per group) were intranasally challenged with PBS (control) or HDM allergen over a 2-week period (on days 0, 7, and 14) to induce allergic inflammation and then exposed to FA21 MWCNT (50 μg) on day 15 by intratracheal administration. BALF and lung tissue were collected 24 h after MWCNT administration on day 16 for analysis of pulmonary inflammation (as illustrated in the experimental design in Figure 1 ). Groups comprised of mice treated with FA21, HDM, or FA21 + HDM. Control mice were treated with carrier alone. (A) BALF cell differential counts were determined and expressed as absolute cell number per mouse of lymphocytes (LYM), eosinophils (EOS), macrophages (MAC), and polymorphonuclear neutrophils (NEU). (B) Cell-associated EPO levels in the BALF were determined by colorimetric assay. (C) CD11b + F4/80 - GR1 - Siglec-F + eosinophil numbers present in the BALF were analyzed by flow cytometry. (D) Peribronchial and perivascular inflammation was assessed by histological analysis of lung tissue sections stained with H E (20×). In addition, inflammatory cells infiltrating the airways were examined by microscopic evaluation and imaging of cytospin preparations of BALF cells stained using Hema3. (E) CysLT levels present in BALF determined by ELISA. Results are mean ± SEM ( n = 6), ∗∗ p

    Article Snippet: Flow Cytometric Analysis of Inflammatory Cells in the BALF Bronchoalveolar lavage fluid cells were collected (from 4–6 mice per group) and the pooled cells were FcγR blocked using 2.4G2 antibody (ATCC) and stained with combinations of the following mouse conjugated mAb: allophycocyanin (APC) or FITC anti-CD3, APC/Cy7 anti-CD4, PE anti-CD8a, APC/Cy7 anti-Ly-6G/Ly6C (Gr-1), PE, FITC or Brilliant Violet 421 anti-CD11b, APC or PE anti-F4/80 (all purchased from BioLegend, San Diego, CA, United States).

    Techniques: Mouse Assay, Colorimetric Assay, Flow Cytometry, Cytometry, Staining, Imaging, Enzyme-linked Immunosorbent Assay

    CRACC-CRACC interaction between macrophages and NK cells enhances the cytokine secretion in vitro. A , Ana-1 cells pre-transfected with liposome encapsulated siNeg or siCRACC was subsequently stimulated with Poly I:C. The expression of CRACC mRNA was assayed by quantitative PCR at 3h time point post the Poly I:C stimulation. B , C D , Ana-1 cells pre-transfected with nanoparticle encapsulated siNeg or siCRACC was cultured in absence or presence of NK cells, which were freshly isolated from mice pre-treated by Poly I:C. Then Ana-1 cells cultured with or without NK cells were stimulated with Poly I:C; and D-GalN was made as control. CRACC expression on Ana-1 cells was analyzed by flow cytometry ( B ) and the cytokine (TNF-α and IFN-γ) in the culture supernatant was assayed by ELISA ( C , D ) at 24h time point post the Poly I:C or D-GalN stimulation; Ana-1 cells were identified as F4/80 + cells. E , Ana-1 cells were cocultured with freshly isolated NK cells from mice pre-treated by Poly I:C; and Ana-1 cells alone or NK cells alone were made as control. The Ana-1-NK coculture was stimulated by Poly I:C in the presence of isotype control Ab (Rat IgG), CRACC mAb (anti-CRACC), or NKG2D mAb (anti-NKG2D), respectively. The IFN-γ in culture supernatant was assayed by ELISA at 24h time point post the Poly I:C stimulation. *P

    Journal: PLoS ONE

    Article Title: CRACC-CRACC Interaction between Kupffer and NK Cells Contributes to Poly I:C/D-GalN Induced Hepatitis

    doi: 10.1371/journal.pone.0076681

    Figure Lengend Snippet: CRACC-CRACC interaction between macrophages and NK cells enhances the cytokine secretion in vitro. A , Ana-1 cells pre-transfected with liposome encapsulated siNeg or siCRACC was subsequently stimulated with Poly I:C. The expression of CRACC mRNA was assayed by quantitative PCR at 3h time point post the Poly I:C stimulation. B , C D , Ana-1 cells pre-transfected with nanoparticle encapsulated siNeg or siCRACC was cultured in absence or presence of NK cells, which were freshly isolated from mice pre-treated by Poly I:C. Then Ana-1 cells cultured with or without NK cells were stimulated with Poly I:C; and D-GalN was made as control. CRACC expression on Ana-1 cells was analyzed by flow cytometry ( B ) and the cytokine (TNF-α and IFN-γ) in the culture supernatant was assayed by ELISA ( C , D ) at 24h time point post the Poly I:C or D-GalN stimulation; Ana-1 cells were identified as F4/80 + cells. E , Ana-1 cells were cocultured with freshly isolated NK cells from mice pre-treated by Poly I:C; and Ana-1 cells alone or NK cells alone were made as control. The Ana-1-NK coculture was stimulated by Poly I:C in the presence of isotype control Ab (Rat IgG), CRACC mAb (anti-CRACC), or NKG2D mAb (anti-NKG2D), respectively. The IFN-γ in culture supernatant was assayed by ELISA at 24h time point post the Poly I:C stimulation. *P

    Article Snippet: Antibodies used for flow cytometry included FITC-conjugated anti-CD3, PE-conjugated anti-NK1.1, PE-conjugated anti-2B4, Percp-Cy5.5-conjugated anti-CD19, Percp-Cy5.5-conjugated anti-F4/80 (BD PharMingen, San Diego), monoclonal anti-mouse CRACC/SLAMF7 antibody which is used combined with a secondary antibody APC-conjugated anti-rat IgG (R & D Systems, Inc.) and Alexa647-conjugated anti-Ly108, Alexa647-conjugated anti-Ly9, Alexa647-conjugated anti-CD84 were kindly provided by Dr. Andrew Veillette (IRCM, CA).

    Techniques: In Vitro, Transfection, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Isolation, Mouse Assay, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    Silencing CRACC expression on Kupffer cells alleviates the liver injury induced by Poly I:C/D-GalN. A - C , Mice were treated with nanoparticle encapsulated FAM conjunct siRNA by intravenous injection; and 3h later FAM + cells among Kupffer cells were analyzed by flow cytometry ( A ); and the nanoparticle encapsulated FAM conjunct siRNA (green) endocytosed by Kupffer cells (red, stained with APC-F4/80 mAb) were tracked by con-focal microscopy both in isolated Kupffer cells ( B , Bar=50µm, 63× objective) and in site of frozen tissue sections( C , Bar=50µm, 40× objective). Kupffer cells were identified as F4/80 + cells. D , Mice were pre-treated with nanoparticle encapsulated siNeg or siCRACC for 6h, and then treated with Poly I:C/D-GalN in each group. The CRACC expression on Kupffer cells was analyzed by flow cytometry at 12h and 18h time points post the Poly I:C/D-GalN treatment. The Data are from 6-8 mice per group. E - F , The mice were pre-treated with nanoparticle encapsulated siNeg or siCRACC for 6h, and then treated with Poly I:C/D-GalN in each group. The serum ALT was tested ( E ) and the H E-Staining analyses were performed at 18h time point post Poly I:C/D-GalN ( F ). The Data are from 6-8 mice per group. **P

    Journal: PLoS ONE

    Article Title: CRACC-CRACC Interaction between Kupffer and NK Cells Contributes to Poly I:C/D-GalN Induced Hepatitis

    doi: 10.1371/journal.pone.0076681

    Figure Lengend Snippet: Silencing CRACC expression on Kupffer cells alleviates the liver injury induced by Poly I:C/D-GalN. A - C , Mice were treated with nanoparticle encapsulated FAM conjunct siRNA by intravenous injection; and 3h later FAM + cells among Kupffer cells were analyzed by flow cytometry ( A ); and the nanoparticle encapsulated FAM conjunct siRNA (green) endocytosed by Kupffer cells (red, stained with APC-F4/80 mAb) were tracked by con-focal microscopy both in isolated Kupffer cells ( B , Bar=50µm, 63× objective) and in site of frozen tissue sections( C , Bar=50µm, 40× objective). Kupffer cells were identified as F4/80 + cells. D , Mice were pre-treated with nanoparticle encapsulated siNeg or siCRACC for 6h, and then treated with Poly I:C/D-GalN in each group. The CRACC expression on Kupffer cells was analyzed by flow cytometry at 12h and 18h time points post the Poly I:C/D-GalN treatment. The Data are from 6-8 mice per group. E - F , The mice were pre-treated with nanoparticle encapsulated siNeg or siCRACC for 6h, and then treated with Poly I:C/D-GalN in each group. The serum ALT was tested ( E ) and the H E-Staining analyses were performed at 18h time point post Poly I:C/D-GalN ( F ). The Data are from 6-8 mice per group. **P

    Article Snippet: Antibodies used for flow cytometry included FITC-conjugated anti-CD3, PE-conjugated anti-NK1.1, PE-conjugated anti-2B4, Percp-Cy5.5-conjugated anti-CD19, Percp-Cy5.5-conjugated anti-F4/80 (BD PharMingen, San Diego), monoclonal anti-mouse CRACC/SLAMF7 antibody which is used combined with a secondary antibody APC-conjugated anti-rat IgG (R & D Systems, Inc.) and Alexa647-conjugated anti-Ly108, Alexa647-conjugated anti-Ly9, Alexa647-conjugated anti-CD84 were kindly provided by Dr. Andrew Veillette (IRCM, CA).

    Techniques: Expressing, Mouse Assay, Injection, Flow Cytometry, Cytometry, Staining, Microscopy, Isolation

    Poly I:C/D-GalN induced liver injury correlates to the increased CRACC mRNA expression of liver. A , Mice were treated with Poly I:C (1.5 µg per mouse, i.v.) together with D-GalN (10 mg per mouse, i.p.); and then euthanized at 2h, 4h, 6h, 8h, 12h, 18h, 24h, 48h time points post Poly I:C/D-GalN treatment. The expression of CRACC mRNA was assayed by quantitative PCR (upper panel); and the serum ALT was tested (lower panel). B , Mice were treated with saline alone, Poly I:C alone, D-GalN alone or Poly I:C together with D-GalN for 12h; and then the CRACC expression on Kupffer cells and hepatic NK cells was analyzed by flow cytometry. Kupffer cells were identified as F4/80 + cells and NK cells were identified as NK1.1 + CD3 - cells. Data are from 6-9 mice per group.

    Journal: PLoS ONE

    Article Title: CRACC-CRACC Interaction between Kupffer and NK Cells Contributes to Poly I:C/D-GalN Induced Hepatitis

    doi: 10.1371/journal.pone.0076681

    Figure Lengend Snippet: Poly I:C/D-GalN induced liver injury correlates to the increased CRACC mRNA expression of liver. A , Mice were treated with Poly I:C (1.5 µg per mouse, i.v.) together with D-GalN (10 mg per mouse, i.p.); and then euthanized at 2h, 4h, 6h, 8h, 12h, 18h, 24h, 48h time points post Poly I:C/D-GalN treatment. The expression of CRACC mRNA was assayed by quantitative PCR (upper panel); and the serum ALT was tested (lower panel). B , Mice were treated with saline alone, Poly I:C alone, D-GalN alone or Poly I:C together with D-GalN for 12h; and then the CRACC expression on Kupffer cells and hepatic NK cells was analyzed by flow cytometry. Kupffer cells were identified as F4/80 + cells and NK cells were identified as NK1.1 + CD3 - cells. Data are from 6-9 mice per group.

    Article Snippet: Antibodies used for flow cytometry included FITC-conjugated anti-CD3, PE-conjugated anti-NK1.1, PE-conjugated anti-2B4, Percp-Cy5.5-conjugated anti-CD19, Percp-Cy5.5-conjugated anti-F4/80 (BD PharMingen, San Diego), monoclonal anti-mouse CRACC/SLAMF7 antibody which is used combined with a secondary antibody APC-conjugated anti-rat IgG (R & D Systems, Inc.) and Alexa647-conjugated anti-Ly108, Alexa647-conjugated anti-Ly9, Alexa647-conjugated anti-CD84 were kindly provided by Dr. Andrew Veillette (IRCM, CA).

    Techniques: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry

    Poly I:C injection elevates the expression of CRACC on Kupffer cells. CRACC expression on alveolar Mϕ, peritoneal Mϕ and Kupffer cells from mice treated with Poly I:C for 18h was analyzed by flow cytometry. Alveolar Mϕ, peritoneal Mϕ and Kupffer cells were identified as F4/80 + cells. A , Flow Cytometry of the CRACC expression on alveolar Mϕ, peritoneal Mϕ and Kupffer cells. B , Percent of CRACC + cells among alveolar Mϕ, peritoneal Mϕ and Kupffer cells. C , Percent of SLAM family (Ly108, Ly9 and CD84) positive cells among Kupffer cells from mice treated with Poly I:C for 18h. Data are from 6-9 mice per group. *P

    Journal: PLoS ONE

    Article Title: CRACC-CRACC Interaction between Kupffer and NK Cells Contributes to Poly I:C/D-GalN Induced Hepatitis

    doi: 10.1371/journal.pone.0076681

    Figure Lengend Snippet: Poly I:C injection elevates the expression of CRACC on Kupffer cells. CRACC expression on alveolar Mϕ, peritoneal Mϕ and Kupffer cells from mice treated with Poly I:C for 18h was analyzed by flow cytometry. Alveolar Mϕ, peritoneal Mϕ and Kupffer cells were identified as F4/80 + cells. A , Flow Cytometry of the CRACC expression on alveolar Mϕ, peritoneal Mϕ and Kupffer cells. B , Percent of CRACC + cells among alveolar Mϕ, peritoneal Mϕ and Kupffer cells. C , Percent of SLAM family (Ly108, Ly9 and CD84) positive cells among Kupffer cells from mice treated with Poly I:C for 18h. Data are from 6-9 mice per group. *P

    Article Snippet: Antibodies used for flow cytometry included FITC-conjugated anti-CD3, PE-conjugated anti-NK1.1, PE-conjugated anti-2B4, Percp-Cy5.5-conjugated anti-CD19, Percp-Cy5.5-conjugated anti-F4/80 (BD PharMingen, San Diego), monoclonal anti-mouse CRACC/SLAMF7 antibody which is used combined with a secondary antibody APC-conjugated anti-rat IgG (R & D Systems, Inc.) and Alexa647-conjugated anti-Ly108, Alexa647-conjugated anti-Ly9, Alexa647-conjugated anti-CD84 were kindly provided by Dr. Andrew Veillette (IRCM, CA).

    Techniques: Injection, Expressing, Mouse Assay, Flow Cytometry, Cytometry

    CRACC expression on lymphocytes and macrophages among various organs. A , Flow Cytometry of the expression of CRACC on lymphocytes (NK cells-solid line, NKT cells-long dashed line, T cells-dashed line and B cells-dotted line) from liver, spleen, lung, peripheral blood (PBL), mesenteric lymph node (MLN) and bone marrow (BM) with mCRACC mAb (open histograms) and isotype control Ab (filled histograms). NK cells were identified as NK1.1 + CD3 - cells; NKT cells were identified as NK1.1 + CD3 + cells; T cells were identified as NK1.1 - CD3 + cells; B cells were identified as CD3 - CD19 + cells. B , Percent of CRACC + cells among lymphocytes from liver, spleen, lung, PBL, MLN and BM. C , Flow cytometry of CRACC expression on macrophages from bronchoalveolar lavage fluid (BALF; alveolar Mϕ), peritoneal cavity (peritoneal Mϕ), and liver (Kupffer cells). Macrophages were identified as F4/80 + cells. D , Percent of CRACC + cells among alveolar Mϕ, peritoneal Mϕ and Kupffer cells. Data are from 6-8 mice per group.

    Journal: PLoS ONE

    Article Title: CRACC-CRACC Interaction between Kupffer and NK Cells Contributes to Poly I:C/D-GalN Induced Hepatitis

    doi: 10.1371/journal.pone.0076681

    Figure Lengend Snippet: CRACC expression on lymphocytes and macrophages among various organs. A , Flow Cytometry of the expression of CRACC on lymphocytes (NK cells-solid line, NKT cells-long dashed line, T cells-dashed line and B cells-dotted line) from liver, spleen, lung, peripheral blood (PBL), mesenteric lymph node (MLN) and bone marrow (BM) with mCRACC mAb (open histograms) and isotype control Ab (filled histograms). NK cells were identified as NK1.1 + CD3 - cells; NKT cells were identified as NK1.1 + CD3 + cells; T cells were identified as NK1.1 - CD3 + cells; B cells were identified as CD3 - CD19 + cells. B , Percent of CRACC + cells among lymphocytes from liver, spleen, lung, PBL, MLN and BM. C , Flow cytometry of CRACC expression on macrophages from bronchoalveolar lavage fluid (BALF; alveolar Mϕ), peritoneal cavity (peritoneal Mϕ), and liver (Kupffer cells). Macrophages were identified as F4/80 + cells. D , Percent of CRACC + cells among alveolar Mϕ, peritoneal Mϕ and Kupffer cells. Data are from 6-8 mice per group.

    Article Snippet: Antibodies used for flow cytometry included FITC-conjugated anti-CD3, PE-conjugated anti-NK1.1, PE-conjugated anti-2B4, Percp-Cy5.5-conjugated anti-CD19, Percp-Cy5.5-conjugated anti-F4/80 (BD PharMingen, San Diego), monoclonal anti-mouse CRACC/SLAMF7 antibody which is used combined with a secondary antibody APC-conjugated anti-rat IgG (R & D Systems, Inc.) and Alexa647-conjugated anti-Ly108, Alexa647-conjugated anti-Ly9, Alexa647-conjugated anti-CD84 were kindly provided by Dr. Andrew Veillette (IRCM, CA).

    Techniques: Expressing, Flow Cytometry, Cytometry, Mouse Assay

    CLEC4F + cells were appeared in the liver environment under Kupffer cell depletion and inflammatory stage. (A) Kupffer cells were depleted by Cl 2 MBP-encapsulated liposome by intravenous injection (100 µl/mouse) at day 0 and livers were harvest at day 1, 7, 14 and 28. F4/80 and CLEC4F immunohistochemistry of liver sections were performed. (B) The numbers of F4/80 + or CLEC4F + cells in livers were shown. For generating inflammatory stage, wild-type and Clec4f−/− littermates were infected with L. monocytogenes (1×10 5 CFU/mouse) intravenously. (C) The numbers of F4/80 + or CLEC4F + cells in livers during L. monocytogenes infection. (D) Immunohistochemistry of L. monocytogenes infected livers of wild-type and Clec4f−/− mice at day 5 after infection. (E) Kaplan-Meier survival curves were shown for Clec4f−/− or wild-type littermates with L. monocytogenes infection. The p value was determined by Log-rank test.

    Journal: PLoS ONE

    Article Title: CLEC4F Is an Inducible C-Type Lectin in F4/80-Positive Cells and Is Involved in Alpha-Galactosylceramide Presentation in Liver

    doi: 10.1371/journal.pone.0065070

    Figure Lengend Snippet: CLEC4F + cells were appeared in the liver environment under Kupffer cell depletion and inflammatory stage. (A) Kupffer cells were depleted by Cl 2 MBP-encapsulated liposome by intravenous injection (100 µl/mouse) at day 0 and livers were harvest at day 1, 7, 14 and 28. F4/80 and CLEC4F immunohistochemistry of liver sections were performed. (B) The numbers of F4/80 + or CLEC4F + cells in livers were shown. For generating inflammatory stage, wild-type and Clec4f−/− littermates were infected with L. monocytogenes (1×10 5 CFU/mouse) intravenously. (C) The numbers of F4/80 + or CLEC4F + cells in livers during L. monocytogenes infection. (D) Immunohistochemistry of L. monocytogenes infected livers of wild-type and Clec4f−/− mice at day 5 after infection. (E) Kaplan-Meier survival curves were shown for Clec4f−/− or wild-type littermates with L. monocytogenes infection. The p value was determined by Log-rank test.

    Article Snippet: Sections were stained with primary antibodies: anti-CLEC4F mAb (clone 3E3F9, 2 µg/ml) and anti-F4/80 polyclonal Ab (M-300, 4 µg/ml, Santa Cruz) followed by HISTOMOUSETM-MAX kit (Zymed) according to the vendor’s instructions.

    Techniques: Injection, Immunohistochemistry, Infection, Mouse Assay

    The expression and distribution of CLEC4F. (A) Tissue distribution of CLEC4F transcripts were analyzed by qRT-PCR. F4/80 is the pan macrophage marker. (B) The protein expression of CLEC4F was examined by Western blot. GAPDH was used as internal control. (C) CLEC4F is a glycoprotein. The pFLAG-CMV-2/CLEC4F-transfected 293T cells were treated with tunicamycin at 1.5 µg/ml for indicated time periods to inhibit N-linked glycosylation, and the molecular weight of CLEC4F was analyzed by Western blot. DMSO is the vehicle control. Mock indicates 293T cells transfect with pFLAG-CMV-2. mLN, mesenteric lymph node; iLN, inguinal lymph node; KC, Kupffer cells; PBL, peripheral blood leukocytes; BM, bone marrow cells; BMDM, bone marrow derived macrophages.

    Journal: PLoS ONE

    Article Title: CLEC4F Is an Inducible C-Type Lectin in F4/80-Positive Cells and Is Involved in Alpha-Galactosylceramide Presentation in Liver

    doi: 10.1371/journal.pone.0065070

    Figure Lengend Snippet: The expression and distribution of CLEC4F. (A) Tissue distribution of CLEC4F transcripts were analyzed by qRT-PCR. F4/80 is the pan macrophage marker. (B) The protein expression of CLEC4F was examined by Western blot. GAPDH was used as internal control. (C) CLEC4F is a glycoprotein. The pFLAG-CMV-2/CLEC4F-transfected 293T cells were treated with tunicamycin at 1.5 µg/ml for indicated time periods to inhibit N-linked glycosylation, and the molecular weight of CLEC4F was analyzed by Western blot. DMSO is the vehicle control. Mock indicates 293T cells transfect with pFLAG-CMV-2. mLN, mesenteric lymph node; iLN, inguinal lymph node; KC, Kupffer cells; PBL, peripheral blood leukocytes; BM, bone marrow cells; BMDM, bone marrow derived macrophages.

    Article Snippet: Sections were stained with primary antibodies: anti-CLEC4F mAb (clone 3E3F9, 2 µg/ml) and anti-F4/80 polyclonal Ab (M-300, 4 µg/ml, Santa Cruz) followed by HISTOMOUSETM-MAX kit (Zymed) according to the vendor’s instructions.

    Techniques: Expressing, Quantitative RT-PCR, Marker, Western Blot, Transfection, Molecular Weight, Derivative Assay

    Expression of CLEC4F during embryogenesis. Yolk sac, embryo and fetal liver in various embryonic stages were collected for CLEC4F detection by (A) qRT-PCR and (B) Western blot. Actin was used as internal control. (C) F4/80 and CLEC4F immunohistochemistry of fetal liver from E14.5 and E17.5, respectively.

    Journal: PLoS ONE

    Article Title: CLEC4F Is an Inducible C-Type Lectin in F4/80-Positive Cells and Is Involved in Alpha-Galactosylceramide Presentation in Liver

    doi: 10.1371/journal.pone.0065070

    Figure Lengend Snippet: Expression of CLEC4F during embryogenesis. Yolk sac, embryo and fetal liver in various embryonic stages were collected for CLEC4F detection by (A) qRT-PCR and (B) Western blot. Actin was used as internal control. (C) F4/80 and CLEC4F immunohistochemistry of fetal liver from E14.5 and E17.5, respectively.

    Article Snippet: Sections were stained with primary antibodies: anti-CLEC4F mAb (clone 3E3F9, 2 µg/ml) and anti-F4/80 polyclonal Ab (M-300, 4 µg/ml, Santa Cruz) followed by HISTOMOUSETM-MAX kit (Zymed) according to the vendor’s instructions.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry

    CLEC4F is co-expressed with F4/80 on liver Kupffer cells. (A) CLEC4F and F4/80 immunohistochemistry of parafilm-embedded liver sections from wild-type and Clec4f−/− mice. (B) Double immunofluorescence of CLEC4F and F4/80 in wild-type livers was performed. Nuclei were counterstained with Hoechst 33342. Signals were determined by confocal microscope (magnification 10×63). (C) Coexpression of CLEC4F and F4/80 on Kupffer cells, but not peritoneal macrophages. Cells were double stained with Alexa Fluor 647-conjugated anti-F4/80 and PE-conjugated anti-CLEC4F mAb. Alexa Fluor 647-conjugated rat IgG2b and PE-conjugated mIgG1 were used as isotype controls.

    Journal: PLoS ONE

    Article Title: CLEC4F Is an Inducible C-Type Lectin in F4/80-Positive Cells and Is Involved in Alpha-Galactosylceramide Presentation in Liver

    doi: 10.1371/journal.pone.0065070

    Figure Lengend Snippet: CLEC4F is co-expressed with F4/80 on liver Kupffer cells. (A) CLEC4F and F4/80 immunohistochemistry of parafilm-embedded liver sections from wild-type and Clec4f−/− mice. (B) Double immunofluorescence of CLEC4F and F4/80 in wild-type livers was performed. Nuclei were counterstained with Hoechst 33342. Signals were determined by confocal microscope (magnification 10×63). (C) Coexpression of CLEC4F and F4/80 on Kupffer cells, but not peritoneal macrophages. Cells were double stained with Alexa Fluor 647-conjugated anti-F4/80 and PE-conjugated anti-CLEC4F mAb. Alexa Fluor 647-conjugated rat IgG2b and PE-conjugated mIgG1 were used as isotype controls.

    Article Snippet: Sections were stained with primary antibodies: anti-CLEC4F mAb (clone 3E3F9, 2 µg/ml) and anti-F4/80 polyclonal Ab (M-300, 4 µg/ml, Santa Cruz) followed by HISTOMOUSETM-MAX kit (Zymed) according to the vendor’s instructions.

    Techniques: Immunohistochemistry, Mouse Assay, Immunofluorescence, Microscopy, Staining