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  • 88
    Biorbyt cxcl4 antibody
    <t>CXCL4</t> and CXCL7 promote resistance to DAC in CD34 + and primary CMML specimens. ( A ) Colony formation was inhibited by DAC but restored with the combination of CXCL4 and CXCL7. CD34 + cells were treated with 1 dose of CXCL4, CXCL7, or both (50 ng/ml each) or with vehicle (PBS containing 0.1% BSA) and daily 10-nM doses of DAC for 3 days. After 3 days of in vitro treatment with DAC, cells were plated in methylcellulose and incubated for 12 to 15 days before colonies were counted. Data represent the mean ± SD. Treatment with 10 nM DAC significantly decreased colony formation but failed to do so in the presence of CXCL7 and CXCL4 together. Shown in the 3 panels are the results of 3 independent experiments. Error bars represent the SD. ( B ) CXCL4 and CXCL7 abrogated the effect of DAC on the viability of primary CMML MNCs. CMML MNCs were treated in vitro for 72 hours with 10 nM DAC alone or in the presence of 50 ng/ml CXCL4, CXCL7, or both. Data represent the mean ± SD. Treatment with DAC alone significantly reduced the viability of these cells, but this effect was lost when CXCL4 or CXCL7 was added to the culture. All data represent independent experiments performed in 3 different CMML patients. Error bars represent the SD. * P
    Cxcl4 Antibody, supplied by Biorbyt, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore cxcl4
    Steroid regulation of <t>CXCL4</t> in hESCs. Estrogen, progesterone, progesterone withdrawal (P withdrawal), or cortisol treatment of hESCs revealed that progesterone withdrawal significantly upregulated (a) CXCL4 mRNA concentrations (n = 5) and (b) CXCL4 protein levels detected by in-cell Western blot (n = 4) and quantified by densitometry (c). Green, CXCL4; red, β -tubulin. In (a) and (c), each box represents lower quartile, median, and upper quartile. Whiskers display minimum and maximum value. * P
    Cxcl4, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    RayBiotech cxcl4
    Calcium dependence of platelet exosome secretion and exosome cargo levels in vitro . Each column and error bar depicts the mean ± sem for results with platelets from 6 healthy subjects. Values for GPVI, <t>CXCL4,</t> CXCL7 and HMGB1 were normalized for
    Cxcl4, supplied by RayBiotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems cxcl4 pf4
    Calcium dependence of platelet exosome secretion and exosome cargo levels in vitro . Each column and error bar depicts the mean ± sem for results with platelets from 6 healthy subjects. Values for GPVI, <t>CXCL4,</t> CXCL7 and HMGB1 were normalized for
    Cxcl4 Pf4, supplied by R&D Systems, used in various techniques. Bioz Stars score: 85/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biorbyt cxcl4 antibody fitc
    Calcium dependence of platelet exosome secretion and exosome cargo levels in vitro . Each column and error bar depicts the mean ± sem for results with platelets from 6 healthy subjects. Values for GPVI, <t>CXCL4,</t> CXCL7 and HMGB1 were normalized for
    Cxcl4 Antibody Fitc, supplied by Biorbyt, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cxcl4  (Abcam)
    91
    Abcam cxcl4
    Co-localisation of <t>CXCL4</t> and CXCL7 with CD68-positive cells. (A) Synovial tissue staining of CXCL4 (red) and CD68 (blue) and (B) staining of CXCL7 (red) and CD68 (blue) showed co-localisation of both cytokines with macrophages. Image representative of rheumatoid arthritis synovium (n=10).
    Cxcl4, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems cxcl4
    Functional analysis of CXCL9 in stellate cells. ( A ) CXCL9 mRNA is expressed in HSCs and is significantly increased ( P = .02) after treatment of the cells with IFN- γ . ( B and C ) Stimulation of LX-2 cells with CXCL9 leads to a dose-dependent down-regulation of TGFB ( P = .01) and COLA1A mRNA expression ( P = .001), respectively. The columns represent mean values ± SEM of 3 independent experiments. ( D ) The down-regulation of collagen by CXCL9 in HSCs is confirmed by Western blot with an antibody detecting the immature (175 kilodaltons) and mature (120 kilodaltons) chain of collagen. ( E ) Both splice variants of the CXCL9 receptor CXCR3 are present in LX-2 cells. ( F ) However, stimulation of the CXCR3-B isoform with <t>CXCL4</t> leads only to a modest increase in COLA1A mRNA.
    Cxcl4, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech cxcl4
    Megakaryocytes maintain quiescence of HSCs in vivo ( a,b ) Representative whole-mount images of sternal BM from control ( a ) and <t>Cxcl4</t> -cre;iDTR mice ( b ) after 7 days of DT treatment. Arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs. Mk are distinguished by their size, morphology and CD41 expression. Vascular endothelial cells are stained with antibodies against CD31 and CD144. Scale bars: 50 μm. ( c ) Quantification of Mk per cross section of whole-mount images of transverse-shaved femoral BM of control and Cxcl4 -cre;iDTR mice after 7 days of DT treatment (representative images are shown in Supplementary Fig. 3a ). n = 8 cross sections from four male mice. ( d,e) Number of HSCs per femur in control and Cxcl4 -cre;iDTR mice ( d ) and representative FACS plots ( e ). n = 5 male mice per group. ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in d . (g) Extreme limiting dilution analysis showing the estimated HSC frequency (solid bar) and confidence intervals (dashed lines) in the BM of control or Cxcl4 -cre;iDTR mice after 7 days of DT treatment. n = 4 (control group) and n = 5 ( Cxcl4 -cre;iDTR group) female recipient mice per dilution, except for the 0.2% BM dose (control group n = 5 and Cxcl4 - cre;iDTR group n = 4). ( h,i ) Percentage of proliferating HSCs in the BM of control and Cxcl4 -cre;iDTR mice (as determined by BrdU incorporation) ( h ) and representative FACS plots ( i ). n = 5 male mice per group. ( j ) Q-PCR analysis of cell cycle-related genes within sorted HSCs. n = 4 independent experiments per group. * P
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    Thermo Fisher cxcl4
    Validating CLAD-associated proteins in recipient rat liver allografts. (a) Immunohistochemistry analysis showed that <t>CXCL4</t> detected on POD 60 was significantly expressed in CLAD liver allografts, compared with the control. (b) Validation by Western blot analysis on POD 60 showed that CXCL4, CXCR3, EGFR, JAK2, STAT3, and Collagen IV were significantly overrepresented in all CLAD liver grafts as compared with the control.
    Cxcl4, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Sino Biological cxcl4
    Validating CLAD-associated proteins in recipient rat liver allografts. (a) Immunohistochemistry analysis showed that <t>CXCL4</t> detected on POD 60 was significantly expressed in CLAD liver allografts, compared with the control. (b) Validation by Western blot analysis on POD 60 showed that CXCL4, CXCR3, EGFR, JAK2, STAT3, and Collagen IV were significantly overrepresented in all CLAD liver grafts as compared with the control.
    Cxcl4, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    R&D Systems recombinant cxcl4
    Gli1 + cells in bone marrow fibrosis are transcriptionally distinct from Gli1 + cells in homeostasis. Bigenic Gli1CreER;tdTomato mice were injected with tamoxifen (3x10mg p.o.) at 8 weeks of age, lethally irradiated at 10 days after the last tamoxifen dose and received c-kit enriched hematopoietic stem cells from wildtype littermates expressing either Thrombopoietin-cDNA (ThPO, n=5, 3 males) or control-cDNA (control, n=4, 3 males; both lentiviral SFFV-iGFP vector backbone). Mice were sacrificed at 70 days after transplantation. Gli1 + cells were sort-purified as lin - GFP - dtTomato + and subjected to RNA sequencing. (A) Principal component analysis (PCA). (B) Heatmap representation with hierarchical clustering (C) Significantly directional enriched (median FDR > 0.05) pathways. (D) Fragments per kilobase of exon per million fragments mapped (FPKM) values of <t>CXCL4</t> in control and ThPO group; mean±SEM. (E) Cytokine analysis in serum and cytokine-free medium supernatant of ckit+ cells expressing ThPO or control (lentiviral SFFV-iGFP vector backbone) or Jak2(V617F) or control (both MCSV-IRES-GFP retroviral backbone). n=2. (F) FPKM value of stromal derived factor 1 (SDF 1/ CXCL12) in control and ThPO group; mean±SEM. (G) Representative images of bone marrow from Gli1CreER t2 ;tdTomato mice transplanted with bone marrow from wildtype littermates expressing either ThPO or control-cDNA stained with the neutral lipid staining LipidTOX. Scale bars 50μm. Arrow indicating LipidTOX positive Gli1 + cells. (H) .
    Recombinant Cxcl4, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    R&D Systems mouse anti human pf4 cxcl4
    Gli1 + cells in bone marrow fibrosis are transcriptionally distinct from Gli1 + cells in homeostasis. Bigenic Gli1CreER;tdTomato mice were injected with tamoxifen (3x10mg p.o.) at 8 weeks of age, lethally irradiated at 10 days after the last tamoxifen dose and received c-kit enriched hematopoietic stem cells from wildtype littermates expressing either Thrombopoietin-cDNA (ThPO, n=5, 3 males) or control-cDNA (control, n=4, 3 males; both lentiviral SFFV-iGFP vector backbone). Mice were sacrificed at 70 days after transplantation. Gli1 + cells were sort-purified as lin - GFP - dtTomato + and subjected to RNA sequencing. (A) Principal component analysis (PCA). (B) Heatmap representation with hierarchical clustering (C) Significantly directional enriched (median FDR > 0.05) pathways. (D) Fragments per kilobase of exon per million fragments mapped (FPKM) values of <t>CXCL4</t> in control and ThPO group; mean±SEM. (E) Cytokine analysis in serum and cytokine-free medium supernatant of ckit+ cells expressing ThPO or control (lentiviral SFFV-iGFP vector backbone) or Jak2(V617F) or control (both MCSV-IRES-GFP retroviral backbone). n=2. (F) FPKM value of stromal derived factor 1 (SDF 1/ CXCL12) in control and ThPO group; mean±SEM. (G) Representative images of bone marrow from Gli1CreER t2 ;tdTomato mice transplanted with bone marrow from wildtype littermates expressing either ThPO or control-cDNA stained with the neutral lipid staining LipidTOX. Scale bars 50μm. Arrow indicating LipidTOX positive Gli1 + cells. (H) .
    Mouse Anti Human Pf4 Cxcl4, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human cxcl4 pf4 antibody
    Gli1 + cells in bone marrow fibrosis are transcriptionally distinct from Gli1 + cells in homeostasis. Bigenic Gli1CreER;tdTomato mice were injected with tamoxifen (3x10mg p.o.) at 8 weeks of age, lethally irradiated at 10 days after the last tamoxifen dose and received c-kit enriched hematopoietic stem cells from wildtype littermates expressing either Thrombopoietin-cDNA (ThPO, n=5, 3 males) or control-cDNA (control, n=4, 3 males; both lentiviral SFFV-iGFP vector backbone). Mice were sacrificed at 70 days after transplantation. Gli1 + cells were sort-purified as lin - GFP - dtTomato + and subjected to RNA sequencing. (A) Principal component analysis (PCA). (B) Heatmap representation with hierarchical clustering (C) Significantly directional enriched (median FDR > 0.05) pathways. (D) Fragments per kilobase of exon per million fragments mapped (FPKM) values of <t>CXCL4</t> in control and ThPO group; mean±SEM. (E) Cytokine analysis in serum and cytokine-free medium supernatant of ckit+ cells expressing ThPO or control (lentiviral SFFV-iGFP vector backbone) or Jak2(V617F) or control (both MCSV-IRES-GFP retroviral backbone). n=2. (F) FPKM value of stromal derived factor 1 (SDF 1/ CXCL12) in control and ThPO group; mean±SEM. (G) Representative images of bone marrow from Gli1CreER t2 ;tdTomato mice transplanted with bone marrow from wildtype littermates expressing either ThPO or control-cDNA stained with the neutral lipid staining LipidTOX. Scale bars 50μm. Arrow indicating LipidTOX positive Gli1 + cells. (H) .
    Human Cxcl4 Pf4 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech cxcl4 pf4
    Gli1 + cells in bone marrow fibrosis are transcriptionally distinct from Gli1 + cells in homeostasis. Bigenic Gli1CreER;tdTomato mice were injected with tamoxifen (3x10mg p.o.) at 8 weeks of age, lethally irradiated at 10 days after the last tamoxifen dose and received c-kit enriched hematopoietic stem cells from wildtype littermates expressing either Thrombopoietin-cDNA (ThPO, n=5, 3 males) or control-cDNA (control, n=4, 3 males; both lentiviral SFFV-iGFP vector backbone). Mice were sacrificed at 70 days after transplantation. Gli1 + cells were sort-purified as lin - GFP - dtTomato + and subjected to RNA sequencing. (A) Principal component analysis (PCA). (B) Heatmap representation with hierarchical clustering (C) Significantly directional enriched (median FDR > 0.05) pathways. (D) Fragments per kilobase of exon per million fragments mapped (FPKM) values of <t>CXCL4</t> in control and ThPO group; mean±SEM. (E) Cytokine analysis in serum and cytokine-free medium supernatant of ckit+ cells expressing ThPO or control (lentiviral SFFV-iGFP vector backbone) or Jak2(V617F) or control (both MCSV-IRES-GFP retroviral backbone). n=2. (F) FPKM value of stromal derived factor 1 (SDF 1/ CXCL12) in control and ThPO group; mean±SEM. (G) Representative images of bone marrow from Gli1CreER t2 ;tdTomato mice transplanted with bone marrow from wildtype littermates expressing either ThPO or control-cDNA stained with the neutral lipid staining LipidTOX. Scale bars 50μm. Arrow indicating LipidTOX positive Gli1 + cells. (H) .
    Cxcl4 Pf4, supplied by PeproTech, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    PeproTech human cxcl4
    <t>CXCL4</t> inhibits the early steps of the HIV-1 replication cycle. ( A ) Time dependence of CXCL4 sensitivity as evaluated using the MAGI assay. Enumeration of blue cells after 48 h denotes the completion of a single infectious cycle of HIV-1. To reduce interexperimental
    Human Cxcl4, supplied by PeproTech, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    American Diagnostica anti cxcl4 antibody
    Correlations between the two biomarkers and response to chemotherapy (A) A stacked bar graph shows a significant association between the combined “High SAA/Low <t>CXCL4”</t> biomarker and histologic response to chemotherapy in all osteosarcoma patients. The “High-risk” group was significantly enriched with poor responders. (B) A stacked bar graph shows a significant association between “Low CXCL4” and poor chemotherapy response.
    Anti Cxcl4 Antibody, supplied by American Diagnostica, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech recominbnat cxcl4
    Correlations between the two biomarkers and response to chemotherapy (A) A stacked bar graph shows a significant association between the combined “High SAA/Low <t>CXCL4”</t> biomarker and histologic response to chemotherapy in all osteosarcoma patients. The “High-risk” group was significantly enriched with poor responders. (B) A stacked bar graph shows a significant association between “Low CXCL4” and poor chemotherapy response.
    Recominbnat Cxcl4, supplied by PeproTech, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    USCN Life mouse pf4 cxcl4
    Correlations between the two biomarkers and response to chemotherapy (A) A stacked bar graph shows a significant association between the combined “High SAA/Low <t>CXCL4”</t> biomarker and histologic response to chemotherapy in all osteosarcoma patients. The “High-risk” group was significantly enriched with poor responders. (B) A stacked bar graph shows a significant association between “Low CXCL4” and poor chemotherapy response.
    Mouse Pf4 Cxcl4, supplied by USCN Life, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam human pf4 elisa kit cxcl4
    Correlations between the two biomarkers and response to chemotherapy (A) A stacked bar graph shows a significant association between the combined “High SAA/Low <t>CXCL4”</t> biomarker and histologic response to chemotherapy in all osteosarcoma patients. The “High-risk” group was significantly enriched with poor responders. (B) A stacked bar graph shows a significant association between “Low CXCL4” and poor chemotherapy response.
    Human Pf4 Elisa Kit Cxcl4, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems murine cxcl4
    <t>CXCL4‐induced</t> neutrophil recruitment is dependent on CXCL2 formation. Levels of (A) CXCL2 and (B) number of neutrophils in the lung after intratracheal challenge with CXCL4. (C) Neutrophil accumulation in the lungs of animals treated with vehicle
    Murine Cxcl4, supplied by R&D Systems, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech murine cxcl4
    <t>CXCL4‐induced</t> neutrophil recruitment is dependent on CXCL2 formation. Levels of (A) CXCL2 and (B) number of neutrophils in the lung after intratracheal challenge with CXCL4. (C) Neutrophil accumulation in the lungs of animals treated with vehicle
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    Santa Cruz Biotechnology cxcl4
    Inhibition of <t>Cxcl4</t> reduces colony formation in vitro and Cxcl4 −/− HSCs show a reduction in engraftment in secondary BM transplantation assays. (A) WT BM cells enriched for c-Kit + were transduced with a Cxcl4 -shRNA vector or control, and
    Cxcl4, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech recombinant murine cxcl4
    Megakaryocytes maintain quiescence of HSCs in vivo ( a,b ) Representative whole-mount images of sternal BM from control ( a ) and <t>Cxcl4</t> -cre;iDTR mice ( b ) after 7 days of DT treatment. Arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs. Mk are distinguished by their size, morphology and CD41 expression. Vascular endothelial cells are stained with antibodies against CD31 and CD144. Scale bars: 50 μm. ( c ) Quantification of Mk per cross section of whole-mount images of transverse-shaved femoral BM of control and Cxcl4 -cre;iDTR mice after 7 days of DT treatment (representative images are shown in Supplementary Fig. 3a ). n = 8 cross sections from four male mice. ( d,e) Number of HSCs per femur in control and Cxcl4 -cre;iDTR mice ( d ) and representative FACS plots ( e ). n = 5 male mice per group. ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in d . (g) Extreme limiting dilution analysis showing the estimated HSC frequency (solid bar) and confidence intervals (dashed lines) in the BM of control or Cxcl4 -cre;iDTR mice after 7 days of DT treatment. n = 4 (control group) and n = 5 ( Cxcl4 -cre;iDTR group) female recipient mice per dilution, except for the 0.2% BM dose (control group n = 5 and Cxcl4 - cre;iDTR group n = 4). ( h,i ) Percentage of proliferating HSCs in the BM of control and Cxcl4 -cre;iDTR mice (as determined by BrdU incorporation) ( h ) and representative FACS plots ( i ). n = 5 male mice per group. ( j ) Q-PCR analysis of cell cycle-related genes within sorted HSCs. n = 4 independent experiments per group. * P
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    Image Search Results


    CXCL4 and CXCL7 promote resistance to DAC in CD34 + and primary CMML specimens. ( A ) Colony formation was inhibited by DAC but restored with the combination of CXCL4 and CXCL7. CD34 + cells were treated with 1 dose of CXCL4, CXCL7, or both (50 ng/ml each) or with vehicle (PBS containing 0.1% BSA) and daily 10-nM doses of DAC for 3 days. After 3 days of in vitro treatment with DAC, cells were plated in methylcellulose and incubated for 12 to 15 days before colonies were counted. Data represent the mean ± SD. Treatment with 10 nM DAC significantly decreased colony formation but failed to do so in the presence of CXCL7 and CXCL4 together. Shown in the 3 panels are the results of 3 independent experiments. Error bars represent the SD. ( B ) CXCL4 and CXCL7 abrogated the effect of DAC on the viability of primary CMML MNCs. CMML MNCs were treated in vitro for 72 hours with 10 nM DAC alone or in the presence of 50 ng/ml CXCL4, CXCL7, or both. Data represent the mean ± SD. Treatment with DAC alone significantly reduced the viability of these cells, but this effect was lost when CXCL4 or CXCL7 was added to the culture. All data represent independent experiments performed in 3 different CMML patients. Error bars represent the SD. * P

    Journal: The Journal of Clinical Investigation

    Article Title: Specific molecular signatures predict decitabine response in chronic myelomonocytic leukemia

    doi: 10.1172/JCI78752

    Figure Lengend Snippet: CXCL4 and CXCL7 promote resistance to DAC in CD34 + and primary CMML specimens. ( A ) Colony formation was inhibited by DAC but restored with the combination of CXCL4 and CXCL7. CD34 + cells were treated with 1 dose of CXCL4, CXCL7, or both (50 ng/ml each) or with vehicle (PBS containing 0.1% BSA) and daily 10-nM doses of DAC for 3 days. After 3 days of in vitro treatment with DAC, cells were plated in methylcellulose and incubated for 12 to 15 days before colonies were counted. Data represent the mean ± SD. Treatment with 10 nM DAC significantly decreased colony formation but failed to do so in the presence of CXCL7 and CXCL4 together. Shown in the 3 panels are the results of 3 independent experiments. Error bars represent the SD. ( B ) CXCL4 and CXCL7 abrogated the effect of DAC on the viability of primary CMML MNCs. CMML MNCs were treated in vitro for 72 hours with 10 nM DAC alone or in the presence of 50 ng/ml CXCL4, CXCL7, or both. Data represent the mean ± SD. Treatment with DAC alone significantly reduced the viability of these cells, but this effect was lost when CXCL4 or CXCL7 was added to the culture. All data represent independent experiments performed in 3 different CMML patients. Error bars represent the SD. * P

    Article Snippet: Antigen retrieval was performed in EDTA (1 mM, pH 8.0) for two 15-minute cycles at maximum power in a microwave oven, and slides were then incubated with a CXCL4 antibody (1:300, catalog 500-P05; PeptroTech) or a CXCL7 antibody (1:50, catalog orb13423; Biorbyt).

    Techniques: In Vitro, Incubation

    CXCL4 and CXCL7 are upregulated in the BM of nonresponders. ( A ) qRT-PCR showing validation of overexpression of CXCL4 , CXCL7 , and ITGB3 in nonresponders; each point represents the mean of triplicate wells for each patient sample; the line and error bars indicate the group mean and SD, respectively. ( B ) Pearson’s correlation analysis of expression levels of CXCL7 and CXCL4 by RNA-seq and qRT-PCR. ( C and D ) Representative IHC images for CXCL4 ( C ) and CXCL7 ( D ) in diagnostic BM biopsies in DAC responders and nonresponders. Original magnification, ×40 ( C and D , left panels), ×63 ( C and D , right panels). Representative images from duplicate experiments are shown.

    Journal: The Journal of Clinical Investigation

    Article Title: Specific molecular signatures predict decitabine response in chronic myelomonocytic leukemia

    doi: 10.1172/JCI78752

    Figure Lengend Snippet: CXCL4 and CXCL7 are upregulated in the BM of nonresponders. ( A ) qRT-PCR showing validation of overexpression of CXCL4 , CXCL7 , and ITGB3 in nonresponders; each point represents the mean of triplicate wells for each patient sample; the line and error bars indicate the group mean and SD, respectively. ( B ) Pearson’s correlation analysis of expression levels of CXCL7 and CXCL4 by RNA-seq and qRT-PCR. ( C and D ) Representative IHC images for CXCL4 ( C ) and CXCL7 ( D ) in diagnostic BM biopsies in DAC responders and nonresponders. Original magnification, ×40 ( C and D , left panels), ×63 ( C and D , right panels). Representative images from duplicate experiments are shown.

    Article Snippet: Antigen retrieval was performed in EDTA (1 mM, pH 8.0) for two 15-minute cycles at maximum power in a microwave oven, and slides were then incubated with a CXCL4 antibody (1:300, catalog 500-P05; PeptroTech) or a CXCL7 antibody (1:50, catalog orb13423; Biorbyt).

    Techniques: Quantitative RT-PCR, Over Expression, Expressing, RNA Sequencing Assay, Immunohistochemistry, Diagnostic Assay

    Steroid regulation of CXCL4 in hESCs. Estrogen, progesterone, progesterone withdrawal (P withdrawal), or cortisol treatment of hESCs revealed that progesterone withdrawal significantly upregulated (a) CXCL4 mRNA concentrations (n = 5) and (b) CXCL4 protein levels detected by in-cell Western blot (n = 4) and quantified by densitometry (c). Green, CXCL4; red, β -tubulin. In (a) and (c), each box represents lower quartile, median, and upper quartile. Whiskers display minimum and maximum value. * P

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Steroids Regulate CXCL4 in the Human Endometrium During Menstruation to Enable Efficient Endometrial Repair

    doi: 10.1210/jc.2016-3604

    Figure Lengend Snippet: Steroid regulation of CXCL4 in hESCs. Estrogen, progesterone, progesterone withdrawal (P withdrawal), or cortisol treatment of hESCs revealed that progesterone withdrawal significantly upregulated (a) CXCL4 mRNA concentrations (n = 5) and (b) CXCL4 protein levels detected by in-cell Western blot (n = 4) and quantified by densitometry (c). Green, CXCL4; red, β -tubulin. In (a) and (c), each box represents lower quartile, median, and upper quartile. Whiskers display minimum and maximum value. * P

    Article Snippet: The connecting well had 20 ng/mL CXCL4 [Sigma-Aldrich; half maximal effective concentration for CXCL4 as found by Baltus et al . in 2005 ( )] in RPMI media; RPMI medium alone was used as a negative control.

    Techniques: In-Cell ELISA

    Cortisol-exposed macrophages show increased migration toward CXCL4. (a) Pretreated macrophages [with macrophage colony-stimulating factor (M-CSF) to give an M0 phenotype, granulocyte macrophage colony-stimulating factor (GM-CSF) and interferon (IFN)- γ to produce an M1 type, cortisol to give an M2 phenotype, or estradiol or progesterone] were plated opposite CXCL4 and photographed after 24 hours. (b) Measuring distance traveled (as a percentage of total distance) confirmed that macrophages pretreated with cortisol migrated significantly farther than other cells. n = 5 separate patient samples; * P

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Steroids Regulate CXCL4 in the Human Endometrium During Menstruation to Enable Efficient Endometrial Repair

    doi: 10.1210/jc.2016-3604

    Figure Lengend Snippet: Cortisol-exposed macrophages show increased migration toward CXCL4. (a) Pretreated macrophages [with macrophage colony-stimulating factor (M-CSF) to give an M0 phenotype, granulocyte macrophage colony-stimulating factor (GM-CSF) and interferon (IFN)- γ to produce an M1 type, cortisol to give an M2 phenotype, or estradiol or progesterone] were plated opposite CXCL4 and photographed after 24 hours. (b) Measuring distance traveled (as a percentage of total distance) confirmed that macrophages pretreated with cortisol migrated significantly farther than other cells. n = 5 separate patient samples; * P

    Article Snippet: The connecting well had 20 ng/mL CXCL4 [Sigma-Aldrich; half maximal effective concentration for CXCL4 as found by Baltus et al . in 2005 ( )] in RPMI media; RPMI medium alone was used as a negative control.

    Techniques: Migration

    (a) CXCL4 in whole human endometrial biopsy specimens from across the menstrual cycle reveals maximal expression during the menstrual phase. Each box represents lower quartile, median, and upper quartile. Whiskers display minimum and maximum values. * P

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Steroids Regulate CXCL4 in the Human Endometrium During Menstruation to Enable Efficient Endometrial Repair

    doi: 10.1210/jc.2016-3604

    Figure Lengend Snippet: (a) CXCL4 in whole human endometrial biopsy specimens from across the menstrual cycle reveals maximal expression during the menstrual phase. Each box represents lower quartile, median, and upper quartile. Whiskers display minimum and maximum values. * P

    Article Snippet: The connecting well had 20 ng/mL CXCL4 [Sigma-Aldrich; half maximal effective concentration for CXCL4 as found by Baltus et al . in 2005 ( )] in RPMI media; RPMI medium alone was used as a negative control.

    Techniques: Expressing

    Steroid regulation of CXCL4 in HEECs. Treating HEECs with estrogen, progesterone, progesterone withdrawal (P withdrawal), and cortisol showed that CXCL4 was significantly upregulated by treatment with cortisol at the (a) mRNA level (n = 4) and (b) protein level (n = 4), quantified by densitometry (c). Green, CXCL4; red, β -tubulin. In (a) and (c), each box represents lower quartile, median, and upper quartile. Whiskers display minimum and maximum value. ** P

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Steroids Regulate CXCL4 in the Human Endometrium During Menstruation to Enable Efficient Endometrial Repair

    doi: 10.1210/jc.2016-3604

    Figure Lengend Snippet: Steroid regulation of CXCL4 in HEECs. Treating HEECs with estrogen, progesterone, progesterone withdrawal (P withdrawal), and cortisol showed that CXCL4 was significantly upregulated by treatment with cortisol at the (a) mRNA level (n = 4) and (b) protein level (n = 4), quantified by densitometry (c). Green, CXCL4; red, β -tubulin. In (a) and (c), each box represents lower quartile, median, and upper quartile. Whiskers display minimum and maximum value. ** P

    Article Snippet: The connecting well had 20 ng/mL CXCL4 [Sigma-Aldrich; half maximal effective concentration for CXCL4 as found by Baltus et al . in 2005 ( )] in RPMI media; RPMI medium alone was used as a negative control.

    Techniques:

    (a) CXCL4 in late secretory and menstrual endometrial biopsy specimens from women with objectively measured NMB (

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Steroids Regulate CXCL4 in the Human Endometrium During Menstruation to Enable Efficient Endometrial Repair

    doi: 10.1210/jc.2016-3604

    Figure Lengend Snippet: (a) CXCL4 in late secretory and menstrual endometrial biopsy specimens from women with objectively measured NMB (

    Article Snippet: The connecting well had 20 ng/mL CXCL4 [Sigma-Aldrich; half maximal effective concentration for CXCL4 as found by Baltus et al . in 2005 ( )] in RPMI media; RPMI medium alone was used as a negative control.

    Techniques:

    Calcium dependence of platelet exosome secretion and exosome cargo levels in vitro . Each column and error bar depicts the mean ± sem for results with platelets from 6 healthy subjects. Values for GPVI, CXCL4, CXCL7 and HMGB1 were normalized for

    Journal: The FASEB Journal

    Article Title: Human plasma platelet-derived exosomes: effects of aspirin

    doi: 10.1096/fj.201500150R

    Figure Lengend Snippet: Calcium dependence of platelet exosome secretion and exosome cargo levels in vitro . Each column and error bar depicts the mean ± sem for results with platelets from 6 healthy subjects. Values for GPVI, CXCL4, CXCL7 and HMGB1 were normalized for

    Article Snippet: The sources of ELISA kits for human proteins were the following: GPVI, CD81, and HMGB1 (Cusabio, American Research Products, Waltham, MA, USA); CXCL7 (Abcam, Cambridge, MA, USA); and CXCL4 (platelet factor 4; RayBiotech, Norcross, GA, USA).

    Techniques: In Vitro

    Effects of aspirin consumption on plasma level of platelet-derived exosomes and their cargo levels. Each column and error bar depicts the mean ± sem for results with platelets from 6 healthy subjects. Values for GPVI, CXCL4, CXCL7, and HMGB1 were

    Journal: The FASEB Journal

    Article Title: Human plasma platelet-derived exosomes: effects of aspirin

    doi: 10.1096/fj.201500150R

    Figure Lengend Snippet: Effects of aspirin consumption on plasma level of platelet-derived exosomes and their cargo levels. Each column and error bar depicts the mean ± sem for results with platelets from 6 healthy subjects. Values for GPVI, CXCL4, CXCL7, and HMGB1 were

    Article Snippet: The sources of ELISA kits for human proteins were the following: GPVI, CD81, and HMGB1 (Cusabio, American Research Products, Waltham, MA, USA); CXCL7 (Abcam, Cambridge, MA, USA); and CXCL4 (platelet factor 4; RayBiotech, Norcross, GA, USA).

    Techniques: Derivative Assay

    Effects of aspirin consumption on platelet exosome secretion and exosome cargo levels ex vivo . Each column and error bar depicts the mean ± sem for results with platelets from 6 healthy subjects. Values for GPVI, CXCL4, CXCL7, and HMGB1 were normalized

    Journal: The FASEB Journal

    Article Title: Human plasma platelet-derived exosomes: effects of aspirin

    doi: 10.1096/fj.201500150R

    Figure Lengend Snippet: Effects of aspirin consumption on platelet exosome secretion and exosome cargo levels ex vivo . Each column and error bar depicts the mean ± sem for results with platelets from 6 healthy subjects. Values for GPVI, CXCL4, CXCL7, and HMGB1 were normalized

    Article Snippet: The sources of ELISA kits for human proteins were the following: GPVI, CD81, and HMGB1 (Cusabio, American Research Products, Waltham, MA, USA); CXCL7 (Abcam, Cambridge, MA, USA); and CXCL4 (platelet factor 4; RayBiotech, Norcross, GA, USA).

    Techniques: Ex Vivo

    Co-localisation of CXCL4 and CXCL7 with CD68-positive cells. (A) Synovial tissue staining of CXCL4 (red) and CD68 (blue) and (B) staining of CXCL7 (red) and CD68 (blue) showed co-localisation of both cytokines with macrophages. Image representative of rheumatoid arthritis synovium (n=10).

    Journal: Annals of the Rheumatic Diseases

    Article Title: Expression of chemokines CXCL4 and CXCL7 by synovial macrophages defines an early stage of rheumatoid arthritis

    doi: 10.1136/annrheumdis-2014-206921

    Figure Lengend Snippet: Co-localisation of CXCL4 and CXCL7 with CD68-positive cells. (A) Synovial tissue staining of CXCL4 (red) and CD68 (blue) and (B) staining of CXCL7 (red) and CD68 (blue) showed co-localisation of both cytokines with macrophages. Image representative of rheumatoid arthritis synovium (n=10).

    Article Snippet: Intriguingly there was also a trend towards higher expression in early RA compared with established RA, suggesting an increase in CXCL4 and CXCL7 levels in the early phase of disease in patients whose arthritis persisted versus those whose arthritis resolved.

    Techniques: Staining

    Expression of CXCL4 and CXCL7 in the synovium is predominantly found outside the vasculature. Staining of CXCL4 and CXCL7 in synovial tissue sections was quantified inside and outside of the vasculature, as assessed by co-staining with von Willebrand factor. Expression of both chemokines was significantly elevated outside the vasculature. RA, rheumatoid arthritis. Kruskal–Wallis test, Dunn's post-test; *p

    Journal: Annals of the Rheumatic Diseases

    Article Title: Expression of chemokines CXCL4 and CXCL7 by synovial macrophages defines an early stage of rheumatoid arthritis

    doi: 10.1136/annrheumdis-2014-206921

    Figure Lengend Snippet: Expression of CXCL4 and CXCL7 in the synovium is predominantly found outside the vasculature. Staining of CXCL4 and CXCL7 in synovial tissue sections was quantified inside and outside of the vasculature, as assessed by co-staining with von Willebrand factor. Expression of both chemokines was significantly elevated outside the vasculature. RA, rheumatoid arthritis. Kruskal–Wallis test, Dunn's post-test; *p

    Article Snippet: Intriguingly there was also a trend towards higher expression in early RA compared with established RA, suggesting an increase in CXCL4 and CXCL7 levels in the early phase of disease in patients whose arthritis persisted versus those whose arthritis resolved.

    Techniques: Expressing, Staining

    Immunofluorescence staining of CXLC4 and CXCL7 in synovial tissue sections. (A) Synovial tissue staining of CXCL4 (red), CD68 (blue), CD41 (green) and von Willebrand factor (vWF) (orange). (B) Synovial tissue staining of CXCL7 (red), CD68 (blue), CD41 (green) and vWF (orange). Nuclear counterstain is shown. Images are representative of early rheumatoid arthritis (RA) synovium (n=10). No staining was observed using isotype and concentration-matched negative controls. Images were taken at ×40 magnification. (C) Quantification of CXCL4 and CXCL7 staining, calculated as the number of pixels per μm 2 over 6× 2×2 tile scans at ×40 magnification, in synovial tissue sections from patients with resolving arthritis (n=9), early RA (n=10) and established RA (n=11). Patients with early RA showed a significantly higher level of CXCL4 (p

    Journal: Annals of the Rheumatic Diseases

    Article Title: Expression of chemokines CXCL4 and CXCL7 by synovial macrophages defines an early stage of rheumatoid arthritis

    doi: 10.1136/annrheumdis-2014-206921

    Figure Lengend Snippet: Immunofluorescence staining of CXLC4 and CXCL7 in synovial tissue sections. (A) Synovial tissue staining of CXCL4 (red), CD68 (blue), CD41 (green) and von Willebrand factor (vWF) (orange). (B) Synovial tissue staining of CXCL7 (red), CD68 (blue), CD41 (green) and vWF (orange). Nuclear counterstain is shown. Images are representative of early rheumatoid arthritis (RA) synovium (n=10). No staining was observed using isotype and concentration-matched negative controls. Images were taken at ×40 magnification. (C) Quantification of CXCL4 and CXCL7 staining, calculated as the number of pixels per μm 2 over 6× 2×2 tile scans at ×40 magnification, in synovial tissue sections from patients with resolving arthritis (n=9), early RA (n=10) and established RA (n=11). Patients with early RA showed a significantly higher level of CXCL4 (p

    Article Snippet: Intriguingly there was also a trend towards higher expression in early RA compared with established RA, suggesting an increase in CXCL4 and CXCL7 levels in the early phase of disease in patients whose arthritis persisted versus those whose arthritis resolved.

    Techniques: Immunofluorescence, Staining, Concentration Assay

    Functional analysis of CXCL9 in stellate cells. ( A ) CXCL9 mRNA is expressed in HSCs and is significantly increased ( P = .02) after treatment of the cells with IFN- γ . ( B and C ) Stimulation of LX-2 cells with CXCL9 leads to a dose-dependent down-regulation of TGFB ( P = .01) and COLA1A mRNA expression ( P = .001), respectively. The columns represent mean values ± SEM of 3 independent experiments. ( D ) The down-regulation of collagen by CXCL9 in HSCs is confirmed by Western blot with an antibody detecting the immature (175 kilodaltons) and mature (120 kilodaltons) chain of collagen. ( E ) Both splice variants of the CXCL9 receptor CXCR3 are present in LX-2 cells. ( F ) However, stimulation of the CXCR3-B isoform with CXCL4 leads only to a modest increase in COLA1A mRNA.

    Journal: Gastroenterology

    Article Title: Antifibrotic Effects of CXCL9 and Its Receptor CXCR3 in Livers of Mice and Humans

    doi: 10.1053/j.gastro.2009.03.053

    Figure Lengend Snippet: Functional analysis of CXCL9 in stellate cells. ( A ) CXCL9 mRNA is expressed in HSCs and is significantly increased ( P = .02) after treatment of the cells with IFN- γ . ( B and C ) Stimulation of LX-2 cells with CXCL9 leads to a dose-dependent down-regulation of TGFB ( P = .01) and COLA1A mRNA expression ( P = .001), respectively. The columns represent mean values ± SEM of 3 independent experiments. ( D ) The down-regulation of collagen by CXCL9 in HSCs is confirmed by Western blot with an antibody detecting the immature (175 kilodaltons) and mature (120 kilodaltons) chain of collagen. ( E ) Both splice variants of the CXCL9 receptor CXCR3 are present in LX-2 cells. ( F ) However, stimulation of the CXCR3-B isoform with CXCL4 leads only to a modest increase in COLA1A mRNA.

    Article Snippet: We therefore also stimulated LX-2 cells with CXCL4, a selective ligand for CXCR3-B.

    Techniques: Functional Assay, Expressing, Western Blot

    Megakaryocytes maintain quiescence of HSCs in vivo ( a,b ) Representative whole-mount images of sternal BM from control ( a ) and Cxcl4 -cre;iDTR mice ( b ) after 7 days of DT treatment. Arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs. Mk are distinguished by their size, morphology and CD41 expression. Vascular endothelial cells are stained with antibodies against CD31 and CD144. Scale bars: 50 μm. ( c ) Quantification of Mk per cross section of whole-mount images of transverse-shaved femoral BM of control and Cxcl4 -cre;iDTR mice after 7 days of DT treatment (representative images are shown in Supplementary Fig. 3a ). n = 8 cross sections from four male mice. ( d,e) Number of HSCs per femur in control and Cxcl4 -cre;iDTR mice ( d ) and representative FACS plots ( e ). n = 5 male mice per group. ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in d . (g) Extreme limiting dilution analysis showing the estimated HSC frequency (solid bar) and confidence intervals (dashed lines) in the BM of control or Cxcl4 -cre;iDTR mice after 7 days of DT treatment. n = 4 (control group) and n = 5 ( Cxcl4 -cre;iDTR group) female recipient mice per dilution, except for the 0.2% BM dose (control group n = 5 and Cxcl4 - cre;iDTR group n = 4). ( h,i ) Percentage of proliferating HSCs in the BM of control and Cxcl4 -cre;iDTR mice (as determined by BrdU incorporation) ( h ) and representative FACS plots ( i ). n = 5 male mice per group. ( j ) Q-PCR analysis of cell cycle-related genes within sorted HSCs. n = 4 independent experiments per group. * P

    Journal: Nature medicine

    Article Title: Megakaryocytes regulate hematopoietic stem cell quiescence via Cxcl4 secretion

    doi: 10.1038/nm.3707

    Figure Lengend Snippet: Megakaryocytes maintain quiescence of HSCs in vivo ( a,b ) Representative whole-mount images of sternal BM from control ( a ) and Cxcl4 -cre;iDTR mice ( b ) after 7 days of DT treatment. Arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs. Mk are distinguished by their size, morphology and CD41 expression. Vascular endothelial cells are stained with antibodies against CD31 and CD144. Scale bars: 50 μm. ( c ) Quantification of Mk per cross section of whole-mount images of transverse-shaved femoral BM of control and Cxcl4 -cre;iDTR mice after 7 days of DT treatment (representative images are shown in Supplementary Fig. 3a ). n = 8 cross sections from four male mice. ( d,e) Number of HSCs per femur in control and Cxcl4 -cre;iDTR mice ( d ) and representative FACS plots ( e ). n = 5 male mice per group. ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in d . (g) Extreme limiting dilution analysis showing the estimated HSC frequency (solid bar) and confidence intervals (dashed lines) in the BM of control or Cxcl4 -cre;iDTR mice after 7 days of DT treatment. n = 4 (control group) and n = 5 ( Cxcl4 -cre;iDTR group) female recipient mice per dilution, except for the 0.2% BM dose (control group n = 5 and Cxcl4 - cre;iDTR group n = 4). ( h,i ) Percentage of proliferating HSCs in the BM of control and Cxcl4 -cre;iDTR mice (as determined by BrdU incorporation) ( h ) and representative FACS plots ( i ). n = 5 male mice per group. ( j ) Q-PCR analysis of cell cycle-related genes within sorted HSCs. n = 4 independent experiments per group. * P

    Article Snippet: Author Contributions : I.B., D.L. and S.P. designed the study, performed the majority of the experiments and analyzed data; J.A. and A.B. performed computational modelling and statistical analysis of the data; M.P.L. performed imaging of Cxcl4 in Mk and provided mice; C.S. and Y.K. contributed to the processing and imaging of whole-mount BM tissues; L.S. genotyped mice; M.P provided mice and interpreted data; P.S.F. supervised the study.

    Techniques: In Vivo, Mouse Assay, Expressing, Staining, FACS, Purification, BrdU Incorporation Assay, Polymerase Chain Reaction

    Megakaryocytes control HSC quiescence via Cxcl4 ( a ) Left, gating strategy for sorting of Mk and immunofluorescence image of a Mk sorted accordingly. Right, Q-PCR analysis of HSC quiescence or proliferation-related genes in Mk. n = 3-7 independent experiments. ( b ) Bar graphs shows Cxcl4 mRNA levels in BM cell populations. n = 3 samples per cell type, except for Mk ( n = 6) and total BM ( n = 5). The upper panels show high power images of sorted Mk from C57BL/6 wild-type (WT) and Cxcl4 −/− mice stained with anti-Cxcl4 antibody and DAPI. ( c ) Concentration of Cxcl4 in BMEF of control and Cxcl4 -cre;iDTR mice after DT treatment, determined by ELISA. n = 5 independent samples per group. ( d ) Percentage of proliferating BrdU + HSCs in cultures of Lineage − cells. ( e ) Number of HSCs per femur in male mice treated with PBS ( n = 5) or 0.2 μg and 1 μg Cxcl4 ( n = 4). ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in ( e ). n = 5 (PBS group and 0.2 μg Cxcl4 group) and n = 4 (1 μg Cxcl4 group) female recipient mice. ( g ) Number of HSCs per femur and percentage of proliferating HSCs, in WT and Cxcl4 −/− mice. n = 3 (WT) and n = 5 ( Cxcl4 −/− ) female mice per group. ( h ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in ( g ). n = 5 female recipient mice per group. ( i,j ) Number of HSCs per femur and percentage of proliferating HSCs in the BM of Cxcl4 −/− ( i ) and Cxcl4 -cre; iDTR ( j ) mice injected with either PBS or 1 μg Cxcl4 during seven days. n = 3 ( Cxcl4 −/− ) and n = 4 ( Cxcl4 -cre;iDTR) male mice per group. * P

    Journal: Nature medicine

    Article Title: Megakaryocytes regulate hematopoietic stem cell quiescence via Cxcl4 secretion

    doi: 10.1038/nm.3707

    Figure Lengend Snippet: Megakaryocytes control HSC quiescence via Cxcl4 ( a ) Left, gating strategy for sorting of Mk and immunofluorescence image of a Mk sorted accordingly. Right, Q-PCR analysis of HSC quiescence or proliferation-related genes in Mk. n = 3-7 independent experiments. ( b ) Bar graphs shows Cxcl4 mRNA levels in BM cell populations. n = 3 samples per cell type, except for Mk ( n = 6) and total BM ( n = 5). The upper panels show high power images of sorted Mk from C57BL/6 wild-type (WT) and Cxcl4 −/− mice stained with anti-Cxcl4 antibody and DAPI. ( c ) Concentration of Cxcl4 in BMEF of control and Cxcl4 -cre;iDTR mice after DT treatment, determined by ELISA. n = 5 independent samples per group. ( d ) Percentage of proliferating BrdU + HSCs in cultures of Lineage − cells. ( e ) Number of HSCs per femur in male mice treated with PBS ( n = 5) or 0.2 μg and 1 μg Cxcl4 ( n = 4). ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in ( e ). n = 5 (PBS group and 0.2 μg Cxcl4 group) and n = 4 (1 μg Cxcl4 group) female recipient mice. ( g ) Number of HSCs per femur and percentage of proliferating HSCs, in WT and Cxcl4 −/− mice. n = 3 (WT) and n = 5 ( Cxcl4 −/− ) female mice per group. ( h ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in ( g ). n = 5 female recipient mice per group. ( i,j ) Number of HSCs per femur and percentage of proliferating HSCs in the BM of Cxcl4 −/− ( i ) and Cxcl4 -cre; iDTR ( j ) mice injected with either PBS or 1 μg Cxcl4 during seven days. n = 3 ( Cxcl4 −/− ) and n = 4 ( Cxcl4 -cre;iDTR) male mice per group. * P

    Article Snippet: Author Contributions : I.B., D.L. and S.P. designed the study, performed the majority of the experiments and analyzed data; J.A. and A.B. performed computational modelling and statistical analysis of the data; M.P.L. performed imaging of Cxcl4 in Mk and provided mice; C.S. and Y.K. contributed to the processing and imaging of whole-mount BM tissues; L.S. genotyped mice; M.P provided mice and interpreted data; P.S.F. supervised the study.

    Techniques: Immunofluorescence, Polymerase Chain Reaction, Mouse Assay, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Purification, Injection

    Megakaryocytes regulate HSC quiescence independently from arterioles ( a – c ) Representative whole-mount image of a sternum compartment ( a ) and magnified high power views ( b,c ). Arterioles are identified by CD31 + CD144 + Sca-1 + expression. Yellow arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs and Mk are distinguished by their size, morphology and CD41 expression. ( b,c ) Illustrative images of the measured distances (yellow dashed lines) between HSCs and Mk as well as arterioles in the BM. Scale bars: 100 μm. ( d ) Two dimensional probability distribution of the distances between HSCs and Mk or arterioles in the sternal BM. From left to right: HSC distribution model in wild-type control animals (Actual; n = 260 HSCs), randomized HSC distribution models generated by computational simulations of random HSC localization (Random 1; n = 1000 HSCs) and random HSC and Mk localizations (Random 2; n = 1000 HSCs) and HSC distribution model in Cxcl4 −/− mice ( n = 128 HSCs). ( e,f ) Localization of HSCs relative to arterioles ( e ) and to Mk ( f ) in Cxcl4 −/− and control mice. Two–sample Kolmogorov-Smirnov test: ( e ) P = 0.9865 and ( f ) P

    Journal: Nature medicine

    Article Title: Megakaryocytes regulate hematopoietic stem cell quiescence via Cxcl4 secretion

    doi: 10.1038/nm.3707

    Figure Lengend Snippet: Megakaryocytes regulate HSC quiescence independently from arterioles ( a – c ) Representative whole-mount image of a sternum compartment ( a ) and magnified high power views ( b,c ). Arterioles are identified by CD31 + CD144 + Sca-1 + expression. Yellow arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs and Mk are distinguished by their size, morphology and CD41 expression. ( b,c ) Illustrative images of the measured distances (yellow dashed lines) between HSCs and Mk as well as arterioles in the BM. Scale bars: 100 μm. ( d ) Two dimensional probability distribution of the distances between HSCs and Mk or arterioles in the sternal BM. From left to right: HSC distribution model in wild-type control animals (Actual; n = 260 HSCs), randomized HSC distribution models generated by computational simulations of random HSC localization (Random 1; n = 1000 HSCs) and random HSC and Mk localizations (Random 2; n = 1000 HSCs) and HSC distribution model in Cxcl4 −/− mice ( n = 128 HSCs). ( e,f ) Localization of HSCs relative to arterioles ( e ) and to Mk ( f ) in Cxcl4 −/− and control mice. Two–sample Kolmogorov-Smirnov test: ( e ) P = 0.9865 and ( f ) P

    Article Snippet: Author Contributions : I.B., D.L. and S.P. designed the study, performed the majority of the experiments and analyzed data; J.A. and A.B. performed computational modelling and statistical analysis of the data; M.P.L. performed imaging of Cxcl4 in Mk and provided mice; C.S. and Y.K. contributed to the processing and imaging of whole-mount BM tissues; L.S. genotyped mice; M.P provided mice and interpreted data; P.S.F. supervised the study.

    Techniques: Expressing, Generated, Mouse Assay

    Validating CLAD-associated proteins in recipient rat liver allografts. (a) Immunohistochemistry analysis showed that CXCL4 detected on POD 60 was significantly expressed in CLAD liver allografts, compared with the control. (b) Validation by Western blot analysis on POD 60 showed that CXCL4, CXCR3, EGFR, JAK2, STAT3, and Collagen IV were significantly overrepresented in all CLAD liver grafts as compared with the control.

    Journal: Journal of Immunology Research

    Article Title: CXCL4 Contributes to the Pathogenesis of Chronic Liver Allograft Dysfunction

    doi: 10.1155/2016/9276986

    Figure Lengend Snippet: Validating CLAD-associated proteins in recipient rat liver allografts. (a) Immunohistochemistry analysis showed that CXCL4 detected on POD 60 was significantly expressed in CLAD liver allografts, compared with the control. (b) Validation by Western blot analysis on POD 60 showed that CXCL4, CXCR3, EGFR, JAK2, STAT3, and Collagen IV were significantly overrepresented in all CLAD liver grafts as compared with the control.

    Article Snippet: Quantitative reverse transcription polymerase chain reaction was carried out for CXCL4 with an Assay from Applied Biosystems (Hs00236998_m1).

    Techniques: Immunohistochemistry, Western Blot

    CXCL4 shRNA reduces the expression of fibrogenesis-associated proteins in isolated HSCs. (a) The isolated HSCs were transfected with CXCL4 shRNA for 72 hours. Western blot analysis of HSCs with antibodies against CXCR3, EGFR, JAK2, STAT3, and Collagen IV. (b) Quantitative presentation of these fibrogenesis-associated proteins. The CXCR3, EGFR, JAK2, STAT3, and Collagen IV levels in the CXCL4 shRNA-treated group decreased as a percent of their respective controls (untreated group). The data are presented as the mean ± SD, n = 3 liver grafts per group. ∗ P

    Journal: Journal of Immunology Research

    Article Title: CXCL4 Contributes to the Pathogenesis of Chronic Liver Allograft Dysfunction

    doi: 10.1155/2016/9276986

    Figure Lengend Snippet: CXCL4 shRNA reduces the expression of fibrogenesis-associated proteins in isolated HSCs. (a) The isolated HSCs were transfected with CXCL4 shRNA for 72 hours. Western blot analysis of HSCs with antibodies against CXCR3, EGFR, JAK2, STAT3, and Collagen IV. (b) Quantitative presentation of these fibrogenesis-associated proteins. The CXCR3, EGFR, JAK2, STAT3, and Collagen IV levels in the CXCL4 shRNA-treated group decreased as a percent of their respective controls (untreated group). The data are presented as the mean ± SD, n = 3 liver grafts per group. ∗ P

    Article Snippet: Quantitative reverse transcription polymerase chain reaction was carried out for CXCL4 with an Assay from Applied Biosystems (Hs00236998_m1).

    Techniques: shRNA, Expressing, Isolation, Transfection, Western Blot

    CXCL4 is associated with CLAD in liver transplant patients. (a) The serum concentration of CXCL4 was significantly increased in patients with CLAD (67.3 ± 3.1 ng/mL) compared to healthy controls (25.1 ± 2.6 ng/mL, P = 0.026). (b) CXCL4 mRNA was significantly increased in subjects with CLAD versus individuals without CLAD ( P = 0.019).

    Journal: Journal of Immunology Research

    Article Title: CXCL4 Contributes to the Pathogenesis of Chronic Liver Allograft Dysfunction

    doi: 10.1155/2016/9276986

    Figure Lengend Snippet: CXCL4 is associated with CLAD in liver transplant patients. (a) The serum concentration of CXCL4 was significantly increased in patients with CLAD (67.3 ± 3.1 ng/mL) compared to healthy controls (25.1 ± 2.6 ng/mL, P = 0.026). (b) CXCL4 mRNA was significantly increased in subjects with CLAD versus individuals without CLAD ( P = 0.019).

    Article Snippet: Quantitative reverse transcription polymerase chain reaction was carried out for CXCL4 with an Assay from Applied Biosystems (Hs00236998_m1).

    Techniques: Concentration Assay

    Blockade of CXCL4 protects recipient rat livers from CLAD. For the blockade experiment, either CXCL4mab (1 mg/kg) or physiological saline (control) was delivered to recipient rats (BN-to-Lewis) through the tail vein once every week from POD 5 for 2 months. (a) Survival time was more prolonged among recipient rats in the CXCL4mab-treated group than in the physiological saline group ( P

    Journal: Journal of Immunology Research

    Article Title: CXCL4 Contributes to the Pathogenesis of Chronic Liver Allograft Dysfunction

    doi: 10.1155/2016/9276986

    Figure Lengend Snippet: Blockade of CXCL4 protects recipient rat livers from CLAD. For the blockade experiment, either CXCL4mab (1 mg/kg) or physiological saline (control) was delivered to recipient rats (BN-to-Lewis) through the tail vein once every week from POD 5 for 2 months. (a) Survival time was more prolonged among recipient rats in the CXCL4mab-treated group than in the physiological saline group ( P

    Article Snippet: Quantitative reverse transcription polymerase chain reaction was carried out for CXCL4 with an Assay from Applied Biosystems (Hs00236998_m1).

    Techniques:

    Gli1 + cells in bone marrow fibrosis are transcriptionally distinct from Gli1 + cells in homeostasis. Bigenic Gli1CreER;tdTomato mice were injected with tamoxifen (3x10mg p.o.) at 8 weeks of age, lethally irradiated at 10 days after the last tamoxifen dose and received c-kit enriched hematopoietic stem cells from wildtype littermates expressing either Thrombopoietin-cDNA (ThPO, n=5, 3 males) or control-cDNA (control, n=4, 3 males; both lentiviral SFFV-iGFP vector backbone). Mice were sacrificed at 70 days after transplantation. Gli1 + cells were sort-purified as lin - GFP - dtTomato + and subjected to RNA sequencing. (A) Principal component analysis (PCA). (B) Heatmap representation with hierarchical clustering (C) Significantly directional enriched (median FDR > 0.05) pathways. (D) Fragments per kilobase of exon per million fragments mapped (FPKM) values of CXCL4 in control and ThPO group; mean±SEM. (E) Cytokine analysis in serum and cytokine-free medium supernatant of ckit+ cells expressing ThPO or control (lentiviral SFFV-iGFP vector backbone) or Jak2(V617F) or control (both MCSV-IRES-GFP retroviral backbone). n=2. (F) FPKM value of stromal derived factor 1 (SDF 1/ CXCL12) in control and ThPO group; mean±SEM. (G) Representative images of bone marrow from Gli1CreER t2 ;tdTomato mice transplanted with bone marrow from wildtype littermates expressing either ThPO or control-cDNA stained with the neutral lipid staining LipidTOX. Scale bars 50μm. Arrow indicating LipidTOX positive Gli1 + cells. (H) .

    Journal: Cell stem cell

    Article Title: Gli1+ mesenchymal stromal cells are a key driver of bone marrow fibrosis and an important cellular therapeutic target

    doi: 10.1016/j.stem.2017.03.008

    Figure Lengend Snippet: Gli1 + cells in bone marrow fibrosis are transcriptionally distinct from Gli1 + cells in homeostasis. Bigenic Gli1CreER;tdTomato mice were injected with tamoxifen (3x10mg p.o.) at 8 weeks of age, lethally irradiated at 10 days after the last tamoxifen dose and received c-kit enriched hematopoietic stem cells from wildtype littermates expressing either Thrombopoietin-cDNA (ThPO, n=5, 3 males) or control-cDNA (control, n=4, 3 males; both lentiviral SFFV-iGFP vector backbone). Mice were sacrificed at 70 days after transplantation. Gli1 + cells were sort-purified as lin - GFP - dtTomato + and subjected to RNA sequencing. (A) Principal component analysis (PCA). (B) Heatmap representation with hierarchical clustering (C) Significantly directional enriched (median FDR > 0.05) pathways. (D) Fragments per kilobase of exon per million fragments mapped (FPKM) values of CXCL4 in control and ThPO group; mean±SEM. (E) Cytokine analysis in serum and cytokine-free medium supernatant of ckit+ cells expressing ThPO or control (lentiviral SFFV-iGFP vector backbone) or Jak2(V617F) or control (both MCSV-IRES-GFP retroviral backbone). n=2. (F) FPKM value of stromal derived factor 1 (SDF 1/ CXCL12) in control and ThPO group; mean±SEM. (G) Representative images of bone marrow from Gli1CreER t2 ;tdTomato mice transplanted with bone marrow from wildtype littermates expressing either ThPO or control-cDNA stained with the neutral lipid staining LipidTOX. Scale bars 50μm. Arrow indicating LipidTOX positive Gli1 + cells. (H) .

    Article Snippet: In another set of experiments, solely Gli1+ cells were cultured in the transwell (600,000 cells/well) and recombinant Cxcl4 (95-P4-025, R & D Systems) in DMSO was added at 400ng/ml to the medium in the bottom well versus DMSO alone.

    Techniques: Mouse Assay, Injection, Irradiation, Expressing, Plasmid Preparation, Transplantation Assay, Purification, RNA Sequencing Assay, Derivative Assay, Staining

    Cxcl4 induces migration and myofibroblastic differentiation of Gli1 + cells (A-E) tdTomato-Gli1 + stromal cells co-cultured with c-kit + hematopoietic stem and progenitor cells (HSPCs) expressing either Thrombopoietin-cDNA (ThPO) or control cDNA (control vector=CV) were treated for 24 hours with vehicle or GANT61. CV= control vector, vehicle; CG= control vector and GANT61; TV= ThPO overexpression and vehicle; TG= ThPO overexpression and GANT61. (A) Relative mRNA expression in tdTomato-Gli1 + stromal cells is shown for platelet-factor 4 (Cxcl4), endothelin 1, matrix metalloproteinase 9 (MMP9), Arachidonate 12-lipoxygenase (ALOX12), Peroxisome proliferator-activated receptor gamma (PPARy), B-cell lymphoma gene 2 (Bcl2) and p21. **p

    Journal: Cell stem cell

    Article Title: Gli1+ mesenchymal stromal cells are a key driver of bone marrow fibrosis and an important cellular therapeutic target

    doi: 10.1016/j.stem.2017.03.008

    Figure Lengend Snippet: Cxcl4 induces migration and myofibroblastic differentiation of Gli1 + cells (A-E) tdTomato-Gli1 + stromal cells co-cultured with c-kit + hematopoietic stem and progenitor cells (HSPCs) expressing either Thrombopoietin-cDNA (ThPO) or control cDNA (control vector=CV) were treated for 24 hours with vehicle or GANT61. CV= control vector, vehicle; CG= control vector and GANT61; TV= ThPO overexpression and vehicle; TG= ThPO overexpression and GANT61. (A) Relative mRNA expression in tdTomato-Gli1 + stromal cells is shown for platelet-factor 4 (Cxcl4), endothelin 1, matrix metalloproteinase 9 (MMP9), Arachidonate 12-lipoxygenase (ALOX12), Peroxisome proliferator-activated receptor gamma (PPARy), B-cell lymphoma gene 2 (Bcl2) and p21. **p

    Article Snippet: In another set of experiments, solely Gli1+ cells were cultured in the transwell (600,000 cells/well) and recombinant Cxcl4 (95-P4-025, R & D Systems) in DMSO was added at 400ng/ml to the medium in the bottom well versus DMSO alone.

    Techniques: Migration, Cell Culture, Expressing, Plasmid Preparation, Over Expression

    CXCL4 inhibits the early steps of the HIV-1 replication cycle. ( A ) Time dependence of CXCL4 sensitivity as evaluated using the MAGI assay. Enumeration of blue cells after 48 h denotes the completion of a single infectious cycle of HIV-1. To reduce interexperimental

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Identification of the platelet-derived chemokine CXCL4/PF-4 as a broad-spectrum HIV-1 inhibitor

    doi: 10.1073/pnas.1207314109

    Figure Lengend Snippet: CXCL4 inhibits the early steps of the HIV-1 replication cycle. ( A ) Time dependence of CXCL4 sensitivity as evaluated using the MAGI assay. Enumeration of blue cells after 48 h denotes the completion of a single infectious cycle of HIV-1. To reduce interexperimental

    Article Snippet: Immunomagnetic beads (4 × 104 per tube) covalently linked to a polyclonal antiserum to rabbit IgG (Invitrogen) were incubated with a polyclonal rabbit IgG antibody to human CXCL4 (Peprotech), washed with PBS containing 0.05% (wt/vol) bovine casein and then loaded with recombinant human CXCL4 (2.5 μg per tube); after removing unbound CXCL4 by repeated washings with PBS, chemokine-armed beads were incubated with 0.5 mL of the viral stocks (isolate HIV-1 IIIB, 20–30 ng of p24 Gag protein per tube; isolate 07USLR, 42 ng of p24 per tube).

    Techniques:

    Inhibition of HIV-1 replication by recombinant human CXCL4. ( A ) Dose-dependent inhibition of a prototypic X4 HIV-1 strain (IIIB) in the immortalized human CD4 + T-cell lines MT-2, PM1, and Sup-T1. ( B ) Dose-dependent inhibition of two HIV-1 variants with

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Identification of the platelet-derived chemokine CXCL4/PF-4 as a broad-spectrum HIV-1 inhibitor

    doi: 10.1073/pnas.1207314109

    Figure Lengend Snippet: Inhibition of HIV-1 replication by recombinant human CXCL4. ( A ) Dose-dependent inhibition of a prototypic X4 HIV-1 strain (IIIB) in the immortalized human CD4 + T-cell lines MT-2, PM1, and Sup-T1. ( B ) Dose-dependent inhibition of two HIV-1 variants with

    Article Snippet: Immunomagnetic beads (4 × 104 per tube) covalently linked to a polyclonal antiserum to rabbit IgG (Invitrogen) were incubated with a polyclonal rabbit IgG antibody to human CXCL4 (Peprotech), washed with PBS containing 0.05% (wt/vol) bovine casein and then loaded with recombinant human CXCL4 (2.5 μg per tube); after removing unbound CXCL4 by repeated washings with PBS, chemokine-armed beads were incubated with 0.5 mL of the viral stocks (isolate HIV-1 IIIB, 20–30 ng of p24 Gag protein per tube; isolate 07USLR, 42 ng of p24 per tube).

    Techniques: Inhibition, Recombinant

    CXCL4 directly interacts with the major HIV-1 envelope glycoprotein, gp120. ( A ) Capture of HIV-1 virions (strain IIIB; X4) by CXCL4-armed immunomagnetic beads. Polyclonal goat anti-CXCL4 IgG and irrelevant goat IgG were used at 20 μg/mL; anti-CXCL4

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Identification of the platelet-derived chemokine CXCL4/PF-4 as a broad-spectrum HIV-1 inhibitor

    doi: 10.1073/pnas.1207314109

    Figure Lengend Snippet: CXCL4 directly interacts with the major HIV-1 envelope glycoprotein, gp120. ( A ) Capture of HIV-1 virions (strain IIIB; X4) by CXCL4-armed immunomagnetic beads. Polyclonal goat anti-CXCL4 IgG and irrelevant goat IgG were used at 20 μg/mL; anti-CXCL4

    Article Snippet: Immunomagnetic beads (4 × 104 per tube) covalently linked to a polyclonal antiserum to rabbit IgG (Invitrogen) were incubated with a polyclonal rabbit IgG antibody to human CXCL4 (Peprotech), washed with PBS containing 0.05% (wt/vol) bovine casein and then loaded with recombinant human CXCL4 (2.5 μg per tube); after removing unbound CXCL4 by repeated washings with PBS, chemokine-armed beads were incubated with 0.5 mL of the viral stocks (isolate HIV-1 IIIB, 20–30 ng of p24 Gag protein per tube; isolate 07USLR, 42 ng of p24 per tube).

    Techniques:

    CXCL4 Has a Broad Spectrum of Antiviral Activity Against Primary HIV-1 Isolates.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Identification of the platelet-derived chemokine CXCL4/PF-4 as a broad-spectrum HIV-1 inhibitor

    doi: 10.1073/pnas.1207314109

    Figure Lengend Snippet: CXCL4 Has a Broad Spectrum of Antiviral Activity Against Primary HIV-1 Isolates.

    Article Snippet: Immunomagnetic beads (4 × 104 per tube) covalently linked to a polyclonal antiserum to rabbit IgG (Invitrogen) were incubated with a polyclonal rabbit IgG antibody to human CXCL4 (Peprotech), washed with PBS containing 0.05% (wt/vol) bovine casein and then loaded with recombinant human CXCL4 (2.5 μg per tube); after removing unbound CXCL4 by repeated washings with PBS, chemokine-armed beads were incubated with 0.5 mL of the viral stocks (isolate HIV-1 IIIB, 20–30 ng of p24 Gag protein per tube; isolate 07USLR, 42 ng of p24 per tube).

    Techniques: Activity Assay

    Correlations between the two biomarkers and response to chemotherapy (A) A stacked bar graph shows a significant association between the combined “High SAA/Low CXCL4” biomarker and histologic response to chemotherapy in all osteosarcoma patients. The “High-risk” group was significantly enriched with poor responders. (B) A stacked bar graph shows a significant association between “Low CXCL4” and poor chemotherapy response.

    Journal: Pediatric blood & cancer

    Article Title: The Prognostic Significance of Circulating Serum Amyloid A and CXC Chemokine Ligand 4 in Osteosarcoma

    doi: 10.1002/pbc.26659

    Figure Lengend Snippet: Correlations between the two biomarkers and response to chemotherapy (A) A stacked bar graph shows a significant association between the combined “High SAA/Low CXCL4” biomarker and histologic response to chemotherapy in all osteosarcoma patients. The “High-risk” group was significantly enriched with poor responders. (B) A stacked bar graph shows a significant association between “Low CXCL4” and poor chemotherapy response.

    Article Snippet: In addition, tumor expressions were measured by immunohistochemistry (IHC) with anti-SAA antibody (SAA, Biosource, Camarillo, CA) and anti-CXCL4 antibody (American Diagnostica, Stamford, Conn) as described previously [ , ].

    Techniques: Biomarker Assay

    Kaplan–Meier analysis of overall survival in OS serum samples (A) Patient survival with respect to metastatic status at diagnosis. (B) Survival of “High-risk” cohort consisting of patients with “High SAA” and “Low CXCL4 levels”, compared to “Low-risk” cohort consisting of all others. (C) “High” and “Low-risk” patients stratified by metastatic status at diagnosis. P values correspond to the log rank-test p-value. “Met” and “Non-Met” refer to patients with and without metastatic disease at initial diagnosis, respectively. “High” and “Low” levels refer to circulating levels either higher or lower than the median concentration for each biomarker. All 5-year survival rates are described in Results.

    Journal: Pediatric blood & cancer

    Article Title: The Prognostic Significance of Circulating Serum Amyloid A and CXC Chemokine Ligand 4 in Osteosarcoma

    doi: 10.1002/pbc.26659

    Figure Lengend Snippet: Kaplan–Meier analysis of overall survival in OS serum samples (A) Patient survival with respect to metastatic status at diagnosis. (B) Survival of “High-risk” cohort consisting of patients with “High SAA” and “Low CXCL4 levels”, compared to “Low-risk” cohort consisting of all others. (C) “High” and “Low-risk” patients stratified by metastatic status at diagnosis. P values correspond to the log rank-test p-value. “Met” and “Non-Met” refer to patients with and without metastatic disease at initial diagnosis, respectively. “High” and “Low” levels refer to circulating levels either higher or lower than the median concentration for each biomarker. All 5-year survival rates are described in Results.

    Article Snippet: In addition, tumor expressions were measured by immunohistochemistry (IHC) with anti-SAA antibody (SAA, Biosource, Camarillo, CA) and anti-CXCL4 antibody (American Diagnostica, Stamford, Conn) as described previously [ , ].

    Techniques: Concentration Assay, Biomarker Assay

    Kaplan–Meier analysis of CXCL4 tumor expression (A) Low tumor CXCL4 expression significantly correlated with poor overall survival in osteosarcoma patients. (B) Tumor CXCL4 expression significantly correlated with overall survival after stratification by metastatic status. “CXCL4 −” refers to low expression (i.e. percentage of stained cells

    Journal: Pediatric blood & cancer

    Article Title: The Prognostic Significance of Circulating Serum Amyloid A and CXC Chemokine Ligand 4 in Osteosarcoma

    doi: 10.1002/pbc.26659

    Figure Lengend Snippet: Kaplan–Meier analysis of CXCL4 tumor expression (A) Low tumor CXCL4 expression significantly correlated with poor overall survival in osteosarcoma patients. (B) Tumor CXCL4 expression significantly correlated with overall survival after stratification by metastatic status. “CXCL4 −” refers to low expression (i.e. percentage of stained cells

    Article Snippet: In addition, tumor expressions were measured by immunohistochemistry (IHC) with anti-SAA antibody (SAA, Biosource, Camarillo, CA) and anti-CXCL4 antibody (American Diagnostica, Stamford, Conn) as described previously [ , ].

    Techniques: Expressing, Staining

    CXCL4‐induced neutrophil recruitment is dependent on CXCL2 formation. Levels of (A) CXCL2 and (B) number of neutrophils in the lung after intratracheal challenge with CXCL4. (C) Neutrophil accumulation in the lungs of animals treated with vehicle

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: CXCL4‐induced neutrophil recruitment is dependent on CXCL2 formation. Levels of (A) CXCL2 and (B) number of neutrophils in the lung after intratracheal challenge with CXCL4. (C) Neutrophil accumulation in the lungs of animals treated with vehicle

    Article Snippet: Inserts were placed in wells containing medium alone (control) or medium plus murine CXCL2 (0.1 µg·mL−1 , R & D Systems) or murine CXCL4 (0.1, 0.5 and 1 µg·mL−1 , R & D Systems).

    Techniques:

    CXCL4 regulates pulmonary recruitment of neutrophils in sepsis. (A) Lung MPO levels at 6 h post‐CLP. (B) Number of BALF neutrophils 24 h after CLP induction. (C) Mac‐1 expression on circulating neutrophils 6 h after

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: CXCL4 regulates pulmonary recruitment of neutrophils in sepsis. (A) Lung MPO levels at 6 h post‐CLP. (B) Number of BALF neutrophils 24 h after CLP induction. (C) Mac‐1 expression on circulating neutrophils 6 h after

    Article Snippet: Inserts were placed in wells containing medium alone (control) or medium plus murine CXCL2 (0.1 µg·mL−1 , R & D Systems) or murine CXCL4 (0.1, 0.5 and 1 µg·mL−1 , R & D Systems).

    Techniques: Expressing

    CXCL4 controls CXC chemokine formation in sepsis. Plasma levels of (A) CXCL1 and (B) CXCL2 and lung levels of (C) CXCL1 and (D) CXCL2 determined 24 h after CLP induction. Animals were treated with vehicle, a control antibody (Ctrl ab) or an anti‐CXCL4

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: CXCL4 controls CXC chemokine formation in sepsis. Plasma levels of (A) CXCL1 and (B) CXCL2 and lung levels of (C) CXCL1 and (D) CXCL2 determined 24 h after CLP induction. Animals were treated with vehicle, a control antibody (Ctrl ab) or an anti‐CXCL4

    Article Snippet: Inserts were placed in wells containing medium alone (control) or medium plus murine CXCL2 (0.1 µg·mL−1 , R & D Systems) or murine CXCL4 (0.1, 0.5 and 1 µg·mL−1 , R & D Systems).

    Techniques:

    Proposed model for neutrophil recruitment in septic lung damage mediated by platelet‐derived CXCL4. Abdominal sepsis triggers Rac1 activation and Rac1‐dependent secretion of CXCL4 from platelets. CXCL4 activates alveolar macrophages (AM)

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: Proposed model for neutrophil recruitment in septic lung damage mediated by platelet‐derived CXCL4. Abdominal sepsis triggers Rac1 activation and Rac1‐dependent secretion of CXCL4 from platelets. CXCL4 activates alveolar macrophages (AM)

    Article Snippet: Inserts were placed in wells containing medium alone (control) or medium plus murine CXCL2 (0.1 µg·mL−1 , R & D Systems) or murine CXCL4 (0.1, 0.5 and 1 µg·mL−1 , R & D Systems).

    Techniques: Derivative Assay, Activation Assay

    Rac1 regulates platelet secretion of CXCL4 in sepsis. Animals were treated with vehicle, NSC23766 (5 mg·kg −1 ), a control ab (Ctrl ab) or an anti‐CD42b ab before CLP induction. (A) elisa was used to quantify the levels of

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: Rac1 regulates platelet secretion of CXCL4 in sepsis. Animals were treated with vehicle, NSC23766 (5 mg·kg −1 ), a control ab (Ctrl ab) or an anti‐CD42b ab before CLP induction. (A) elisa was used to quantify the levels of

    Article Snippet: Inserts were placed in wells containing medium alone (control) or medium plus murine CXCL2 (0.1 µg·mL−1 , R & D Systems) or murine CXCL4 (0.1, 0.5 and 1 µg·mL−1 , R & D Systems).

    Techniques: Enzyme-linked Immunosorbent Assay

    Rac1 regulates agonist‐induced secretion of CXCL4 in platelets. (A) Isolated platelets were incubated with or without NSC23766 (10 μM) and then stimulated with recombinant PAR4 (200 μM), and the level of CXCL4 in

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: Rac1 regulates agonist‐induced secretion of CXCL4 in platelets. (A) Isolated platelets were incubated with or without NSC23766 (10 μM) and then stimulated with recombinant PAR4 (200 μM), and the level of CXCL4 in

    Article Snippet: Inserts were placed in wells containing medium alone (control) or medium plus murine CXCL2 (0.1 µg·mL−1 , R & D Systems) or murine CXCL4 (0.1, 0.5 and 1 µg·mL−1 , R & D Systems).

    Techniques: Isolation, Incubation, Recombinant

    CXCL4 regulates lung damage in sepsis. (A) Oedema formation in the lung. (B) Representative haematoxylin and eosin sections of lung are shown. Animals were treated with vehicle, a control ab (Ctrl ab) or an anti‐CXCL4 ab before CLP induction.

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: CXCL4 regulates lung damage in sepsis. (A) Oedema formation in the lung. (B) Representative haematoxylin and eosin sections of lung are shown. Animals were treated with vehicle, a control ab (Ctrl ab) or an anti‐CXCL4 ab before CLP induction.

    Article Snippet: Inserts were placed in wells containing medium alone (control) or medium plus murine CXCL2 (0.1 µg·mL−1 , R & D Systems) or murine CXCL4 (0.1, 0.5 and 1 µg·mL−1 , R & D Systems).

    Techniques:

    CXCL4‐induced neutrophil recruitment is dependent on CXCL2 formation. Levels of (A) CXCL2 and (B) number of neutrophils in the lung after intratracheal challenge with CXCL4. (C) Neutrophil accumulation in the lungs of animals treated with vehicle

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: CXCL4‐induced neutrophil recruitment is dependent on CXCL2 formation. Levels of (A) CXCL2 and (B) number of neutrophils in the lung after intratracheal challenge with CXCL4. (C) Neutrophil accumulation in the lungs of animals treated with vehicle

    Article Snippet: Intratracheal administration of CXCL4 was performed via a skin incision over the trachea and injection of murine CXCL4 (1 µg, PeproTech Nordic, Stockholm, Sweden) into the trachea with a 27G needle in anaesthetized animals.

    Techniques:

    CXCL4 regulates pulmonary recruitment of neutrophils in sepsis. (A) Lung MPO levels at 6 h post‐CLP. (B) Number of BALF neutrophils 24 h after CLP induction. (C) Mac‐1 expression on circulating neutrophils 6 h after

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: CXCL4 regulates pulmonary recruitment of neutrophils in sepsis. (A) Lung MPO levels at 6 h post‐CLP. (B) Number of BALF neutrophils 24 h after CLP induction. (C) Mac‐1 expression on circulating neutrophils 6 h after

    Article Snippet: Intratracheal administration of CXCL4 was performed via a skin incision over the trachea and injection of murine CXCL4 (1 µg, PeproTech Nordic, Stockholm, Sweden) into the trachea with a 27G needle in anaesthetized animals.

    Techniques: Expressing

    CXCL4 controls CXC chemokine formation in sepsis. Plasma levels of (A) CXCL1 and (B) CXCL2 and lung levels of (C) CXCL1 and (D) CXCL2 determined 24 h after CLP induction. Animals were treated with vehicle, a control antibody (Ctrl ab) or an anti‐CXCL4

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: CXCL4 controls CXC chemokine formation in sepsis. Plasma levels of (A) CXCL1 and (B) CXCL2 and lung levels of (C) CXCL1 and (D) CXCL2 determined 24 h after CLP induction. Animals were treated with vehicle, a control antibody (Ctrl ab) or an anti‐CXCL4

    Article Snippet: Intratracheal administration of CXCL4 was performed via a skin incision over the trachea and injection of murine CXCL4 (1 µg, PeproTech Nordic, Stockholm, Sweden) into the trachea with a 27G needle in anaesthetized animals.

    Techniques:

    Proposed model for neutrophil recruitment in septic lung damage mediated by platelet‐derived CXCL4. Abdominal sepsis triggers Rac1 activation and Rac1‐dependent secretion of CXCL4 from platelets. CXCL4 activates alveolar macrophages (AM)

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: Proposed model for neutrophil recruitment in septic lung damage mediated by platelet‐derived CXCL4. Abdominal sepsis triggers Rac1 activation and Rac1‐dependent secretion of CXCL4 from platelets. CXCL4 activates alveolar macrophages (AM)

    Article Snippet: Intratracheal administration of CXCL4 was performed via a skin incision over the trachea and injection of murine CXCL4 (1 µg, PeproTech Nordic, Stockholm, Sweden) into the trachea with a 27G needle in anaesthetized animals.

    Techniques: Derivative Assay, Activation Assay

    Rac1 regulates platelet secretion of CXCL4 in sepsis. Animals were treated with vehicle, NSC23766 (5 mg·kg −1 ), a control ab (Ctrl ab) or an anti‐CD42b ab before CLP induction. (A) elisa was used to quantify the levels of

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: Rac1 regulates platelet secretion of CXCL4 in sepsis. Animals were treated with vehicle, NSC23766 (5 mg·kg −1 ), a control ab (Ctrl ab) or an anti‐CD42b ab before CLP induction. (A) elisa was used to quantify the levels of

    Article Snippet: Intratracheal administration of CXCL4 was performed via a skin incision over the trachea and injection of murine CXCL4 (1 µg, PeproTech Nordic, Stockholm, Sweden) into the trachea with a 27G needle in anaesthetized animals.

    Techniques: Enzyme-linked Immunosorbent Assay

    Rac1 regulates agonist‐induced secretion of CXCL4 in platelets. (A) Isolated platelets were incubated with or without NSC23766 (10 μM) and then stimulated with recombinant PAR4 (200 μM), and the level of CXCL4 in

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: Rac1 regulates agonist‐induced secretion of CXCL4 in platelets. (A) Isolated platelets were incubated with or without NSC23766 (10 μM) and then stimulated with recombinant PAR4 (200 μM), and the level of CXCL4 in

    Article Snippet: Intratracheal administration of CXCL4 was performed via a skin incision over the trachea and injection of murine CXCL4 (1 µg, PeproTech Nordic, Stockholm, Sweden) into the trachea with a 27G needle in anaesthetized animals.

    Techniques: Isolation, Incubation, Recombinant

    CXCL4 regulates lung damage in sepsis. (A) Oedema formation in the lung. (B) Representative haematoxylin and eosin sections of lung are shown. Animals were treated with vehicle, a control ab (Ctrl ab) or an anti‐CXCL4 ab before CLP induction.

    Journal: British Journal of Pharmacology

    Article Title: Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage) Platelet secretion of CXCL4 is Rac1‐dependent and regulates neutrophil infiltration and tissue damage in septic lung damage

    doi: 10.1111/bph.13325

    Figure Lengend Snippet: CXCL4 regulates lung damage in sepsis. (A) Oedema formation in the lung. (B) Representative haematoxylin and eosin sections of lung are shown. Animals were treated with vehicle, a control ab (Ctrl ab) or an anti‐CXCL4 ab before CLP induction.

    Article Snippet: Intratracheal administration of CXCL4 was performed via a skin incision over the trachea and injection of murine CXCL4 (1 µg, PeproTech Nordic, Stockholm, Sweden) into the trachea with a 27G needle in anaesthetized animals.

    Techniques:

    Inhibition of Cxcl4 reduces colony formation in vitro and Cxcl4 −/− HSCs show a reduction in engraftment in secondary BM transplantation assays. (A) WT BM cells enriched for c-Kit + were transduced with a Cxcl4 -shRNA vector or control, and

    Journal: Blood

    Article Title: CXCR2 and CXCL4 regulate survival and self-renewal of hematopoietic stem/progenitor cells

    doi: 10.1182/blood-2015-08-661785

    Figure Lengend Snippet: Inhibition of Cxcl4 reduces colony formation in vitro and Cxcl4 −/− HSCs show a reduction in engraftment in secondary BM transplantation assays. (A) WT BM cells enriched for c-Kit + were transduced with a Cxcl4 -shRNA vector or control, and

    Article Snippet: Cells were fixed and permeabilized with 4% formaldehyde and 0.25% Triton X-100, blocked in 10% goat or donkey serum (Sigma-Aldrich) and incubated overnight in primary antibodies against CXCL1 (1:100, sc-1374; Santa Cruz, Heidelberg, Germany), CXCL2 (1:100, sc-1375; Santa Cruz), CXCL4 (1:100, sc-73638; Santa Cruz), CXCL6 (1:100, sc-5813; Santa Cruz), CXCL8 (1:100, MAB208; R & D Systems, Abingdon, United Kingdom), CXCR2 (1:100, sc-682; Santa Cruz), or immunoglobulin G (AB-108-C; R & D Systems).

    Techniques: Inhibition, In Vitro, Transplantation Assay, Transduction, shRNA, Plasmid Preparation

    CXCL4 and CXCR2 are expressed on CD34 + 38 − and CD34 + 38 + cells. (A) RT-PCR in human CD34 + 38 − and CD34 + 38 + cells shows gene expression differences for the chemokine ligands CXCL1 , CXCL2 , CXCL4 , CXCL6 , and CXCL8 . Fold change was calculated

    Journal: Blood

    Article Title: CXCR2 and CXCL4 regulate survival and self-renewal of hematopoietic stem/progenitor cells

    doi: 10.1182/blood-2015-08-661785

    Figure Lengend Snippet: CXCL4 and CXCR2 are expressed on CD34 + 38 − and CD34 + 38 + cells. (A) RT-PCR in human CD34 + 38 − and CD34 + 38 + cells shows gene expression differences for the chemokine ligands CXCL1 , CXCL2 , CXCL4 , CXCL6 , and CXCL8 . Fold change was calculated

    Article Snippet: Cells were fixed and permeabilized with 4% formaldehyde and 0.25% Triton X-100, blocked in 10% goat or donkey serum (Sigma-Aldrich) and incubated overnight in primary antibodies against CXCL1 (1:100, sc-1374; Santa Cruz, Heidelberg, Germany), CXCL2 (1:100, sc-1375; Santa Cruz), CXCL4 (1:100, sc-73638; Santa Cruz), CXCL6 (1:100, sc-5813; Santa Cruz), CXCL8 (1:100, MAB208; R & D Systems, Abingdon, United Kingdom), CXCR2 (1:100, sc-682; Santa Cruz), or immunoglobulin G (AB-108-C; R & D Systems).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

    Inhibition of CXCL4 ligands and CXCR2 signaling reduces cell viability and colony formation in vitro. (A) CD34 + enriched cells were transduced with CXCL4 siRNA, and relative scrambled control and level of knock-down (left) and viability (right) measured

    Journal: Blood

    Article Title: CXCR2 and CXCL4 regulate survival and self-renewal of hematopoietic stem/progenitor cells

    doi: 10.1182/blood-2015-08-661785

    Figure Lengend Snippet: Inhibition of CXCL4 ligands and CXCR2 signaling reduces cell viability and colony formation in vitro. (A) CD34 + enriched cells were transduced with CXCL4 siRNA, and relative scrambled control and level of knock-down (left) and viability (right) measured

    Article Snippet: Cells were fixed and permeabilized with 4% formaldehyde and 0.25% Triton X-100, blocked in 10% goat or donkey serum (Sigma-Aldrich) and incubated overnight in primary antibodies against CXCL1 (1:100, sc-1374; Santa Cruz, Heidelberg, Germany), CXCL2 (1:100, sc-1375; Santa Cruz), CXCL4 (1:100, sc-73638; Santa Cruz), CXCL6 (1:100, sc-5813; Santa Cruz), CXCL8 (1:100, MAB208; R & D Systems, Abingdon, United Kingdom), CXCR2 (1:100, sc-682; Santa Cruz), or immunoglobulin G (AB-108-C; R & D Systems).

    Techniques: Inhibition, In Vitro, Transduction

    Cxcl4 −/− mice display a reduction in HSC compartment. (A) BM (left) and spleen (right) from WT mice (n = 6) or Cxcl4 −/− mice (n = 6) were harvested and total cellularity assessed. (B) Numbers of LSK cells for BM (left)

    Journal: Blood

    Article Title: CXCR2 and CXCL4 regulate survival and self-renewal of hematopoietic stem/progenitor cells

    doi: 10.1182/blood-2015-08-661785

    Figure Lengend Snippet: Cxcl4 −/− mice display a reduction in HSC compartment. (A) BM (left) and spleen (right) from WT mice (n = 6) or Cxcl4 −/− mice (n = 6) were harvested and total cellularity assessed. (B) Numbers of LSK cells for BM (left)

    Article Snippet: Cells were fixed and permeabilized with 4% formaldehyde and 0.25% Triton X-100, blocked in 10% goat or donkey serum (Sigma-Aldrich) and incubated overnight in primary antibodies against CXCL1 (1:100, sc-1374; Santa Cruz, Heidelberg, Germany), CXCL2 (1:100, sc-1375; Santa Cruz), CXCL4 (1:100, sc-73638; Santa Cruz), CXCL6 (1:100, sc-5813; Santa Cruz), CXCL8 (1:100, MAB208; R & D Systems, Abingdon, United Kingdom), CXCR2 (1:100, sc-682; Santa Cruz), or immunoglobulin G (AB-108-C; R & D Systems).

    Techniques: Mouse Assay

    Megakaryocytes maintain quiescence of HSCs in vivo ( a,b ) Representative whole-mount images of sternal BM from control ( a ) and Cxcl4 -cre;iDTR mice ( b ) after 7 days of DT treatment. Arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs. Mk are distinguished by their size, morphology and CD41 expression. Vascular endothelial cells are stained with antibodies against CD31 and CD144. Scale bars: 50 μm. ( c ) Quantification of Mk per cross section of whole-mount images of transverse-shaved femoral BM of control and Cxcl4 -cre;iDTR mice after 7 days of DT treatment (representative images are shown in Supplementary Fig. 3a ). n = 8 cross sections from four male mice. ( d,e) Number of HSCs per femur in control and Cxcl4 -cre;iDTR mice ( d ) and representative FACS plots ( e ). n = 5 male mice per group. ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in d . (g) Extreme limiting dilution analysis showing the estimated HSC frequency (solid bar) and confidence intervals (dashed lines) in the BM of control or Cxcl4 -cre;iDTR mice after 7 days of DT treatment. n = 4 (control group) and n = 5 ( Cxcl4 -cre;iDTR group) female recipient mice per dilution, except for the 0.2% BM dose (control group n = 5 and Cxcl4 - cre;iDTR group n = 4). ( h,i ) Percentage of proliferating HSCs in the BM of control and Cxcl4 -cre;iDTR mice (as determined by BrdU incorporation) ( h ) and representative FACS plots ( i ). n = 5 male mice per group. ( j ) Q-PCR analysis of cell cycle-related genes within sorted HSCs. n = 4 independent experiments per group. * P

    Journal: Nature medicine

    Article Title: Megakaryocytes regulate hematopoietic stem cell quiescence via Cxcl4 secretion

    doi: 10.1038/nm.3707

    Figure Lengend Snippet: Megakaryocytes maintain quiescence of HSCs in vivo ( a,b ) Representative whole-mount images of sternal BM from control ( a ) and Cxcl4 -cre;iDTR mice ( b ) after 7 days of DT treatment. Arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs. Mk are distinguished by their size, morphology and CD41 expression. Vascular endothelial cells are stained with antibodies against CD31 and CD144. Scale bars: 50 μm. ( c ) Quantification of Mk per cross section of whole-mount images of transverse-shaved femoral BM of control and Cxcl4 -cre;iDTR mice after 7 days of DT treatment (representative images are shown in Supplementary Fig. 3a ). n = 8 cross sections from four male mice. ( d,e) Number of HSCs per femur in control and Cxcl4 -cre;iDTR mice ( d ) and representative FACS plots ( e ). n = 5 male mice per group. ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in d . (g) Extreme limiting dilution analysis showing the estimated HSC frequency (solid bar) and confidence intervals (dashed lines) in the BM of control or Cxcl4 -cre;iDTR mice after 7 days of DT treatment. n = 4 (control group) and n = 5 ( Cxcl4 -cre;iDTR group) female recipient mice per dilution, except for the 0.2% BM dose (control group n = 5 and Cxcl4 - cre;iDTR group n = 4). ( h,i ) Percentage of proliferating HSCs in the BM of control and Cxcl4 -cre;iDTR mice (as determined by BrdU incorporation) ( h ) and representative FACS plots ( i ). n = 5 male mice per group. ( j ) Q-PCR analysis of cell cycle-related genes within sorted HSCs. n = 4 independent experiments per group. * P

    Article Snippet: Recombinant murine Cxcl4 (100 ng ml−1 ; PeproTech) or vehicle with or without heparin (30 μg ml−1 ; Sigma) was added to assess Cxcl4-related effects on HSCs.

    Techniques: In Vivo, Mouse Assay, Expressing, Staining, FACS, Purification, BrdU Incorporation Assay, Polymerase Chain Reaction

    Megakaryocytes control HSC quiescence via Cxcl4 ( a ) Left, gating strategy for sorting of Mk and immunofluorescence image of a Mk sorted accordingly. Right, Q-PCR analysis of HSC quiescence or proliferation-related genes in Mk. n = 3-7 independent experiments. ( b ) Bar graphs shows Cxcl4 mRNA levels in BM cell populations. n = 3 samples per cell type, except for Mk ( n = 6) and total BM ( n = 5). The upper panels show high power images of sorted Mk from C57BL/6 wild-type (WT) and Cxcl4 −/− mice stained with anti-Cxcl4 antibody and DAPI. ( c ) Concentration of Cxcl4 in BMEF of control and Cxcl4 -cre;iDTR mice after DT treatment, determined by ELISA. n = 5 independent samples per group. ( d ) Percentage of proliferating BrdU + HSCs in cultures of Lineage − cells. ( e ) Number of HSCs per femur in male mice treated with PBS ( n = 5) or 0.2 μg and 1 μg Cxcl4 ( n = 4). ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in ( e ). n = 5 (PBS group and 0.2 μg Cxcl4 group) and n = 4 (1 μg Cxcl4 group) female recipient mice. ( g ) Number of HSCs per femur and percentage of proliferating HSCs, in WT and Cxcl4 −/− mice. n = 3 (WT) and n = 5 ( Cxcl4 −/− ) female mice per group. ( h ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in ( g ). n = 5 female recipient mice per group. ( i,j ) Number of HSCs per femur and percentage of proliferating HSCs in the BM of Cxcl4 −/− ( i ) and Cxcl4 -cre; iDTR ( j ) mice injected with either PBS or 1 μg Cxcl4 during seven days. n = 3 ( Cxcl4 −/− ) and n = 4 ( Cxcl4 -cre;iDTR) male mice per group. * P

    Journal: Nature medicine

    Article Title: Megakaryocytes regulate hematopoietic stem cell quiescence via Cxcl4 secretion

    doi: 10.1038/nm.3707

    Figure Lengend Snippet: Megakaryocytes control HSC quiescence via Cxcl4 ( a ) Left, gating strategy for sorting of Mk and immunofluorescence image of a Mk sorted accordingly. Right, Q-PCR analysis of HSC quiescence or proliferation-related genes in Mk. n = 3-7 independent experiments. ( b ) Bar graphs shows Cxcl4 mRNA levels in BM cell populations. n = 3 samples per cell type, except for Mk ( n = 6) and total BM ( n = 5). The upper panels show high power images of sorted Mk from C57BL/6 wild-type (WT) and Cxcl4 −/− mice stained with anti-Cxcl4 antibody and DAPI. ( c ) Concentration of Cxcl4 in BMEF of control and Cxcl4 -cre;iDTR mice after DT treatment, determined by ELISA. n = 5 independent samples per group. ( d ) Percentage of proliferating BrdU + HSCs in cultures of Lineage − cells. ( e ) Number of HSCs per femur in male mice treated with PBS ( n = 5) or 0.2 μg and 1 μg Cxcl4 ( n = 4). ( f ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in ( e ). n = 5 (PBS group and 0.2 μg Cxcl4 group) and n = 4 (1 μg Cxcl4 group) female recipient mice. ( g ) Number of HSCs per femur and percentage of proliferating HSCs, in WT and Cxcl4 −/− mice. n = 3 (WT) and n = 5 ( Cxcl4 −/− ) female mice per group. ( h ) Percentage of CD45.2 + cells in the blood of CD45.1 + mice competitively transplanted with total BM cells purified from the mice analysed in ( g ). n = 5 female recipient mice per group. ( i,j ) Number of HSCs per femur and percentage of proliferating HSCs in the BM of Cxcl4 −/− ( i ) and Cxcl4 -cre; iDTR ( j ) mice injected with either PBS or 1 μg Cxcl4 during seven days. n = 3 ( Cxcl4 −/− ) and n = 4 ( Cxcl4 -cre;iDTR) male mice per group. * P

    Article Snippet: Recombinant murine Cxcl4 (100 ng ml−1 ; PeproTech) or vehicle with or without heparin (30 μg ml−1 ; Sigma) was added to assess Cxcl4-related effects on HSCs.

    Techniques: Immunofluorescence, Polymerase Chain Reaction, Mouse Assay, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Purification, Injection

    Megakaryocytes regulate HSC quiescence independently from arterioles ( a – c ) Representative whole-mount image of a sternum compartment ( a ) and magnified high power views ( b,c ). Arterioles are identified by CD31 + CD144 + Sca-1 + expression. Yellow arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs and Mk are distinguished by their size, morphology and CD41 expression. ( b,c ) Illustrative images of the measured distances (yellow dashed lines) between HSCs and Mk as well as arterioles in the BM. Scale bars: 100 μm. ( d ) Two dimensional probability distribution of the distances between HSCs and Mk or arterioles in the sternal BM. From left to right: HSC distribution model in wild-type control animals (Actual; n = 260 HSCs), randomized HSC distribution models generated by computational simulations of random HSC localization (Random 1; n = 1000 HSCs) and random HSC and Mk localizations (Random 2; n = 1000 HSCs) and HSC distribution model in Cxcl4 −/− mice ( n = 128 HSCs). ( e,f ) Localization of HSCs relative to arterioles ( e ) and to Mk ( f ) in Cxcl4 −/− and control mice. Two–sample Kolmogorov-Smirnov test: ( e ) P = 0.9865 and ( f ) P

    Journal: Nature medicine

    Article Title: Megakaryocytes regulate hematopoietic stem cell quiescence via Cxcl4 secretion

    doi: 10.1038/nm.3707

    Figure Lengend Snippet: Megakaryocytes regulate HSC quiescence independently from arterioles ( a – c ) Representative whole-mount image of a sternum compartment ( a ) and magnified high power views ( b,c ). Arterioles are identified by CD31 + CD144 + Sca-1 + expression. Yellow arrowheads denote Lin − CD48 − CD41 − CD150 + phenotypic HSCs and Mk are distinguished by their size, morphology and CD41 expression. ( b,c ) Illustrative images of the measured distances (yellow dashed lines) between HSCs and Mk as well as arterioles in the BM. Scale bars: 100 μm. ( d ) Two dimensional probability distribution of the distances between HSCs and Mk or arterioles in the sternal BM. From left to right: HSC distribution model in wild-type control animals (Actual; n = 260 HSCs), randomized HSC distribution models generated by computational simulations of random HSC localization (Random 1; n = 1000 HSCs) and random HSC and Mk localizations (Random 2; n = 1000 HSCs) and HSC distribution model in Cxcl4 −/− mice ( n = 128 HSCs). ( e,f ) Localization of HSCs relative to arterioles ( e ) and to Mk ( f ) in Cxcl4 −/− and control mice. Two–sample Kolmogorov-Smirnov test: ( e ) P = 0.9865 and ( f ) P

    Article Snippet: Recombinant murine Cxcl4 (100 ng ml−1 ; PeproTech) or vehicle with or without heparin (30 μg ml−1 ; Sigma) was added to assess Cxcl4-related effects on HSCs.

    Techniques: Expressing, Generated, Mouse Assay