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  • 99
    Thermo Fisher gene exp retnla mm00445109 m1
    Gene Exp Retnla Mm00445109 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 220 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Abcam fizz 1 antibody
    The pleiotropic effects of MTD chemotherapy–treated CAFs mediated through the ELR + chemokine−CXCR-2 paracrine signaling axis. (A) Bar graphs showing the increased endothelial tube formation induced by MTD-doxorubicin– or MTD-paclitaxel–treated BCAF-011 CAFs that could be reduced by treatment with the CXCR-2 inhibitor SB225002 (1 µM) or prevented when LDM-mimetic regimens as described in Fig. 5 (A and C) were used to treat CAFs. Data from two independent experiments ( n = 3 in each group) are shown. (B, left) Representative photomicrographs showing pronounced vascularity in tumors formed by orthotopically implanted BC-011 carcinoma cells along with MTD-doxorubicin (MTD-CAF)– or vehicle (vehicle-CAF)-treated CAFs with or without concurrent intraperitoneal injections of SB225002 (0.5 mg/kg/day) in NOG mice. Also shown are immunohistochemical images of von Willebrand factor (vWF) staining of the endothelial cells in the tumors. Bar, 50 µm. (Right) quantification of microvessel density (MVD) per high-power field (HPF) in tumors. At least 10 different areas were examined in each tissue. Data from one experiment ( n = 3 in each group) are shown. (C, left) The invasive capacities of U937-derived macrophages in response to vehicle-CAFs, MTD-CAFs, or LDM-CAFs in the absence or presence of 1 µM SB225002 in a Transwell invasion assay. Shown are representative immunofluorescence images of the invaded cells, with cell nuclei stained with CYTOX-green. Bars, 100 µm. (Right) The numbers of invaded macrophages. Data from two independent experiments ( n = 3 in each group) are shown. (D, left) Representative immunohistochemical images of F4/80 staining in tumors formed by subcutaneous inoculation of BC-011 cells along with vehicle-, MTD-, or LDM-CAFs in the flank of nude mice. Bar, 50 µm. (Right) Percent F4/80- or <t>Fizz-1–positive</t> cells in the microscopic field. At least 10 different areas were examined in each tissue. Data from two independent experiment ( n = 3 tumor tissues in each group) are shown. (E) BCAF-011 CAFs were treated with vehicle (vehicle-CAF), MTD, (MTD-CAF), or LDM-doxorubicin or paclitaxel (LDM-CAF) as in Fig. 5 and then co-cultivated with BC-011 carcinoma cells in the presence or absence of 1 µM SB225002 in a dual chamber culture apparatus for 5 d, after which the carcinoma cells were subjected to flow cytometric analyses. Shown are the percentages of the CD44 + CD24 −/low carcinoma cells. Data from two independent experiments ( n = 3 in each group) are shown. (F) Freshly sorted CD44 + CD24 −/low BC-011 carcinoma cells were cultured in the presence of human recombinant CXCL-1, -2, -5, and -6 (each at 1 µg/ml) or vehicle in low-attachment culture plates for 10 d, and the diameters of the tumorspheres generated were quantified. Data from two independent experiments ( n = 3 in each group) are shown. (G) The ability of CD44 + CD24 −/low BC-011 carcinoma cells to invade through reconstituted basement membrane in response to human recombinant CXCL-1, -2, -5, and -6 (each at 1 µg/ml) in a Transwell invasion assay. Data from two independent experiments ( n = 3 in each group) are shown. (H) The percentages of CD44 + CD24 −/low cells in BC-011 carcinoma cells co-cultivated with vehicle- or MTD-CAFs along with the neutralizing antibody directed against individual ELR + chemokines. Data from two independent experiments ( n = 3 in each group) are shown. (I) Limiting dilution assay demonstrating the tumorsphere formation efficiency of BC-011 carcinoma cells cultured in the conditioned medium from MTD- or LDM-CAFs. The arrow indicates change of slope of the trend line, suggestive of differential tumorsphere formation ability. Data from one experiment ( n = 6 in each group; mean ± SEM; Student’s t test; ***, P
    Fizz 1 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    PeproTech recombinant fizz 1
    The pleiotropic effects of MTD chemotherapy–treated CAFs mediated through the ELR + chemokine−CXCR-2 paracrine signaling axis. (A) Bar graphs showing the increased endothelial tube formation induced by MTD-doxorubicin– or MTD-paclitaxel–treated BCAF-011 CAFs that could be reduced by treatment with the CXCR-2 inhibitor SB225002 (1 µM) or prevented when LDM-mimetic regimens as described in Fig. 5 (A and C) were used to treat CAFs. Data from two independent experiments ( n = 3 in each group) are shown. (B, left) Representative photomicrographs showing pronounced vascularity in tumors formed by orthotopically implanted BC-011 carcinoma cells along with MTD-doxorubicin (MTD-CAF)– or vehicle (vehicle-CAF)-treated CAFs with or without concurrent intraperitoneal injections of SB225002 (0.5 mg/kg/day) in NOG mice. Also shown are immunohistochemical images of von Willebrand factor (vWF) staining of the endothelial cells in the tumors. Bar, 50 µm. (Right) quantification of microvessel density (MVD) per high-power field (HPF) in tumors. At least 10 different areas were examined in each tissue. Data from one experiment ( n = 3 in each group) are shown. (C, left) The invasive capacities of U937-derived macrophages in response to vehicle-CAFs, MTD-CAFs, or LDM-CAFs in the absence or presence of 1 µM SB225002 in a Transwell invasion assay. Shown are representative immunofluorescence images of the invaded cells, with cell nuclei stained with CYTOX-green. Bars, 100 µm. (Right) The numbers of invaded macrophages. Data from two independent experiments ( n = 3 in each group) are shown. (D, left) Representative immunohistochemical images of F4/80 staining in tumors formed by subcutaneous inoculation of BC-011 cells along with vehicle-, MTD-, or LDM-CAFs in the flank of nude mice. Bar, 50 µm. (Right) Percent F4/80- or <t>Fizz-1–positive</t> cells in the microscopic field. At least 10 different areas were examined in each tissue. Data from two independent experiment ( n = 3 tumor tissues in each group) are shown. (E) BCAF-011 CAFs were treated with vehicle (vehicle-CAF), MTD, (MTD-CAF), or LDM-doxorubicin or paclitaxel (LDM-CAF) as in Fig. 5 and then co-cultivated with BC-011 carcinoma cells in the presence or absence of 1 µM SB225002 in a dual chamber culture apparatus for 5 d, after which the carcinoma cells were subjected to flow cytometric analyses. Shown are the percentages of the CD44 + CD24 −/low carcinoma cells. Data from two independent experiments ( n = 3 in each group) are shown. (F) Freshly sorted CD44 + CD24 −/low BC-011 carcinoma cells were cultured in the presence of human recombinant CXCL-1, -2, -5, and -6 (each at 1 µg/ml) or vehicle in low-attachment culture plates for 10 d, and the diameters of the tumorspheres generated were quantified. Data from two independent experiments ( n = 3 in each group) are shown. (G) The ability of CD44 + CD24 −/low BC-011 carcinoma cells to invade through reconstituted basement membrane in response to human recombinant CXCL-1, -2, -5, and -6 (each at 1 µg/ml) in a Transwell invasion assay. Data from two independent experiments ( n = 3 in each group) are shown. (H) The percentages of CD44 + CD24 −/low cells in BC-011 carcinoma cells co-cultivated with vehicle- or MTD-CAFs along with the neutralizing antibody directed against individual ELR + chemokines. Data from two independent experiments ( n = 3 in each group) are shown. (I) Limiting dilution assay demonstrating the tumorsphere formation efficiency of BC-011 carcinoma cells cultured in the conditioned medium from MTD- or LDM-CAFs. The arrow indicates change of slope of the trend line, suggestive of differential tumorsphere formation ability. Data from one experiment ( n = 6 in each group; mean ± SEM; Student’s t test; ***, P
    Recombinant Fizz 1, supplied by PeproTech, used in various techniques. Bioz Stars score: 81/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    fizz1  (Abcam)
    87
    Abcam fizz1
    DHT enhances M2 gene expression, YM1 and <t>FIZZ1</t> production in BMM. Bone marrow from female and male C57BL/6 mice was cultured for 10 days with M-CSF (40 ng/mL) to obtain BMM. (A) Validation of the specificity of anti-AR antibody. AM from female and male mice were cultured for 24 h in the presence or absence of the AR-degrading drug, ASC-J9 (10 μM), left panel. AR expression in BMM from C57BL/6 female and male mice, middle panel. Comparison of AR expression in BMM and AM, right panel. (B) BMM were cultured for 10 days with 40 ng/mL of GM-CSF or M-CSF in 6-well plates, cells were collected and viability determined. F4/80 + cells were selected in the live population. Dot plots of CD11c + Siglec F + cells from F4/80 + cells cultured with GM-CSF or M-CSF. (C) Analysis of double-positive CD11c + Siglec F + macrophages with GM-CSF or M-CSF (D) Analysis of AR + cells from F4/80 + cells cultured with GM-CSF or M-CSF. (E) BMM were pretreated overnight with the indicated concentrations of DHT and then stimulated with IL-4 (1 ng/mL) for 48 h. Expression of the indicated M2 genes was determined by qPCR using the 2 -ΔΔCT method, and compared to the amount of mRNA in the male IL-4 sample (= 100%). (F) Expression of YM1 and FIZZ1 protein was determined by Western blot in the same BMM supernatants after 48 h of IL-4 stimulation. Equal volumes of supernatant were loaded in each lane. Densitometry of the specific bands was normalized to the amount of protein in male IL-4 (= 100%). Results are representative of four independent in vitro experiments. † p
    Fizz1, supplied by Abcam, used in various techniques. Bioz Stars score: 87/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    R&D Systems fizz1
    <t>FIZZ1</t> induces myofibroblast differentiation. A: Fully differentiated 3T3-L1 adipocytes were treated with 25 ng/mL FIZZ1 for 7 days. The indicated genes were analyzed by RT-qPCR. B: α-SMA protein induction with indicated doses of FIZZ1 was analyzed
    Fizz1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 89/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    Alpha Diagnostics fizz1
    Histological examination of lung tissue. Tissues from pre-terminal bronchioles were analyzed for morphologic features, mucosubstances in mucous cells and the presence of some proteins associated with allergic lung inflammation. (A-C) H E staining. (D-E) AB/PAS staining. Arrows depict AB/PAS-stained mucosubstances in airway epithelium. (G-I) Immunohistological staining using anti Ym1/Ym2 antibodies. Arrows depict stained Ym1/Ym2. (J-L) Immunohistological staining using anti <t>FIZZ1</t> antibodies. Arrows depict FIZZ in the epithelium. Abbreviations: AB/PAS, Alcian blue-Periodic acid schiff double stain; AL, airway lumen; ap, alveolar parenchyma; bv, blood vessel; e, airway surface epithelium; H E, hematoxylin and eosin stain.
    Fizz1, supplied by Alpha Diagnostics, used in various techniques. Bioz Stars score: 81/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    83
    PeproTech fizz1
    Lung macrophages prominently express M2 proteins following IL-4-treatment in intratracheal LPS-exposed WT mice. A : mRNA expression for the M2 markers Arg1 , <t>Fizz1</t> , and Ym1 quantified in the whole lung at days 4 and 6 among intratracheal H 2 O- or LPS-exposed
    Fizz1, supplied by PeproTech, used in various techniques. Bioz Stars score: 83/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    83
    Santa Cruz Biotechnology fizz1
    STAT6 signaling pathway involvement in gefitinib-mediated inhibition of M2-like polarization of macrophages. (A) RAW 264.7 cells were treated with IL-13 (10 ng/mL) with or without gefitinib for the indicated times. Western blots of STAT6 and activated p-STAT6 in cell lysates. (B) Western blot was performed to measure the protein level of Mrc1, Arg1, Ym1, and <t>Fizz1.</t> BMDMs were stimulated with IL-13 (10 ng/mL) and treated with or without gefitinib for 24 h. Actin was used as a loading control. The results are presented as the mean±SEM of triplicate determinations. * P
    Fizz1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 83/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    GeneTex inflammatory zone 1 fizz 1
    STAT6 signaling pathway involvement in gefitinib-mediated inhibition of M2-like polarization of macrophages. (A) RAW 264.7 cells were treated with IL-13 (10 ng/mL) with or without gefitinib for the indicated times. Western blots of STAT6 and activated p-STAT6 in cell lysates. (B) Western blot was performed to measure the protein level of Mrc1, Arg1, Ym1, and <t>Fizz1.</t> BMDMs were stimulated with IL-13 (10 ng/mL) and treated with or without gefitinib for 24 h. Actin was used as a loading control. The results are presented as the mean±SEM of triplicate determinations. * P
    Inflammatory Zone 1 Fizz 1, supplied by GeneTex, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    R&D Systems fizz1 relm α
    STAT6 signaling pathway involvement in gefitinib-mediated inhibition of M2-like polarization of macrophages. (A) RAW 264.7 cells were treated with IL-13 (10 ng/mL) with or without gefitinib for the indicated times. Western blots of STAT6 and activated p-STAT6 in cell lysates. (B) Western blot was performed to measure the protein level of Mrc1, Arg1, Ym1, and <t>Fizz1.</t> BMDMs were stimulated with IL-13 (10 ng/mL) and treated with or without gefitinib for 24 h. Actin was used as a loading control. The results are presented as the mean±SEM of triplicate determinations. * P
    Fizz1 Relm α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Santa Cruz Biotechnology fizz1 recombinant protein
    STAT6 signaling pathway involvement in gefitinib-mediated inhibition of M2-like polarization of macrophages. (A) RAW 264.7 cells were treated with IL-13 (10 ng/mL) with or without gefitinib for the indicated times. Western blots of STAT6 and activated p-STAT6 in cell lysates. (B) Western blot was performed to measure the protein level of Mrc1, Arg1, Ym1, and <t>Fizz1.</t> BMDMs were stimulated with IL-13 (10 ng/mL) and treated with or without gefitinib for 24 h. Actin was used as a loading control. The results are presented as the mean±SEM of triplicate determinations. * P
    Fizz1 Recombinant Protein, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Abcam relmα fizz1
    Switch to M2a polarization in IKKβ cKO mice at mid-infection stage. M2a activation occurs in IKKβ cKO mice after sublethal i.d. infection with 10 6 Ft . LVS as evidenced by expression of ( A ) CD206 ( B ) IL-10 ( C ) <t>FIZZ1/Relmα</t> and ( D ) Arg-1 in a flow cytometry time course experiment. Statistical analysis was performed by one-way ANOVA followed by Tukey’s ad hoc post-test. (n for IKK f/f , IKKα cKO, IKKβ cKO on day 0∶5/5/5; day 2∶4/4/4; day 6∶4/4/4; day 8∶4/4/3). Results are representative of at least two independent experiments. Bars represent the mean ± SD. **P
    Relmα Fizz1, supplied by Abcam, used in various techniques. Bioz Stars score: 84/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    R&D Systems mouse fizz1 relm alpha antibody
    Switch to M2a polarization in IKKβ cKO mice at mid-infection stage. M2a activation occurs in IKKβ cKO mice after sublethal i.d. infection with 10 6 Ft . LVS as evidenced by expression of ( A ) CD206 ( B ) IL-10 ( C ) <t>FIZZ1/Relmα</t> and ( D ) Arg-1 in a flow cytometry time course experiment. Statistical analysis was performed by one-way ANOVA followed by Tukey’s ad hoc post-test. (n for IKK f/f , IKKα cKO, IKKβ cKO on day 0∶5/5/5; day 2∶4/4/4; day 6∶4/4/4; day 8∶4/4/3). Results are representative of at least two independent experiments. Bars represent the mean ± SD. **P
    Mouse Fizz1 Relm Alpha Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    83
    PeproTech rabbit polyclonal anti fizz 1
    Switch to M2a polarization in IKKβ cKO mice at mid-infection stage. M2a activation occurs in IKKβ cKO mice after sublethal i.d. infection with 10 6 Ft . LVS as evidenced by expression of ( A ) CD206 ( B ) IL-10 ( C ) <t>FIZZ1/Relmα</t> and ( D ) Arg-1 in a flow cytometry time course experiment. Statistical analysis was performed by one-way ANOVA followed by Tukey’s ad hoc post-test. (n for IKK f/f , IKKα cKO, IKKβ cKO on day 0∶5/5/5; day 2∶4/4/4; day 6∶4/4/4; day 8∶4/4/3). Results are representative of at least two independent experiments. Bars represent the mean ± SD. **P
    Rabbit Polyclonal Anti Fizz 1, supplied by PeproTech, used in various techniques. Bioz Stars score: 83/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    R&D Systems rat anti fizz1 relmα antibody
    Switch to M2a polarization in IKKβ cKO mice at mid-infection stage. M2a activation occurs in IKKβ cKO mice after sublethal i.d. infection with 10 6 Ft . LVS as evidenced by expression of ( A ) CD206 ( B ) IL-10 ( C ) <t>FIZZ1/Relmα</t> and ( D ) Arg-1 in a flow cytometry time course experiment. Statistical analysis was performed by one-way ANOVA followed by Tukey’s ad hoc post-test. (n for IKK f/f , IKKα cKO, IKKβ cKO on day 0∶5/5/5; day 2∶4/4/4; day 6∶4/4/4; day 8∶4/4/3). Results are representative of at least two independent experiments. Bars represent the mean ± SD. **P
    Rat Anti Fizz1 Relmα Antibody, supplied by R&D Systems, 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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    80
    R&D Systems mouse fizz1 antibody
    Moderate dose of XBJ (5 mg/mL) enhanced the expression of M2 phenotype markers of mouse peritoneal macrophages. Peritoneal macrophages of mice were, respectively, treated with 0 mg/mL, 1 mg/mL, 5 mg/mL, 10 mg/mL, 50 mg/mL, and 100 mg/mL XBJ in the presence of 100 ng/mL LPS for 6 h, 24 h, or 48 h. Cytokines levels including TNF- α , IL-6, and IL-10 in the medium were determined by ELISA (a). F4/80, CD11c, and CD206 expressions were determined by flow cytometry. The group of F4/80 + CD11c + CD206 − was classified as M1 and F4/80 + CD11c − CD206 + as M2 (b). The treated macrophages were lysed and levels of Arg1, Ym1, and <t>Fizz1</t> were measured by western blot (c). Data represented the mean ± SEM of independent experiment in triplicate. ∗ p
    Mouse Fizz1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 80/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Becton Dickinson anti mouse fizz1
    Moderate dose of XBJ (5 mg/mL) enhanced the expression of M2 phenotype markers of mouse peritoneal macrophages. Peritoneal macrophages of mice were, respectively, treated with 0 mg/mL, 1 mg/mL, 5 mg/mL, 10 mg/mL, 50 mg/mL, and 100 mg/mL XBJ in the presence of 100 ng/mL LPS for 6 h, 24 h, or 48 h. Cytokines levels including TNF- α , IL-6, and IL-10 in the medium were determined by ELISA (a). F4/80, CD11c, and CD206 expressions were determined by flow cytometry. The group of F4/80 + CD11c + CD206 − was classified as M1 and F4/80 + CD11c − CD206 + as M2 (b). The treated macrophages were lysed and levels of Arg1, Ym1, and <t>Fizz1</t> were measured by western blot (c). Data represented the mean ± SEM of independent experiment in triplicate. ∗ p
    Anti Mouse Fizz1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 80/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    Enzo Biochem anti rat fizz1
    The lung HYP content was measured in <t>GFP-FIZZ1</t> chimera lung samples after BLM or PBS treatment. Data were shown as percentages of their respective PBS controls. The mean values were shown from 2 mice each group. *P
    Anti Rat Fizz1, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 81/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    R&D Systems recombinant mouse fizz1
    The lung HYP content was measured in <t>GFP-FIZZ1</t> chimera lung samples after BLM or PBS treatment. Data were shown as percentages of their respective PBS controls. The mean values were shown from 2 mice each group. *P
    Recombinant Mouse Fizz1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Leinco Technologies recombinant mouse fizz1
    Effect of <t>FIZZ1</t> treatment on lung fibroblast-expressed Notch1 signaling components. Normal murine lung fibroblasts were treated with the indicated doses of rmFIZZ1 for 1 hour, or as indicated. A : Cell lysates (20 μg protein each) were analyzed
    Recombinant Mouse Fizz1, supplied by Leinco Technologies, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Abcam m2 marker fizz1
    Effect of <t>FIZZ1</t> treatment on lung fibroblast-expressed Notch1 signaling components. Normal murine lung fibroblasts were treated with the indicated doses of rmFIZZ1 for 1 hour, or as indicated. A : Cell lysates (20 μg protein each) were analyzed
    M2 Marker Fizz1, supplied by Abcam, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    Abcam rabbit polyclonal fizz1 antibody
    Effect of <t>FIZZ1</t> treatment on lung fibroblast-expressed Notch1 signaling components. Normal murine lung fibroblasts were treated with the indicated doses of rmFIZZ1 for 1 hour, or as indicated. A : Cell lysates (20 μg protein each) were analyzed
    Rabbit Polyclonal Fizz1 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 82/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Photomicrographs of M2 microglia following GCM stimulation of glioma cells in vitro and in vivo. (a) The human microglia HMC3 cells were treated with GCM from LN229, LN229R, HG7 and HG7R cells. (b) HMC3 were treated with GCM from LN229-Scr, LN229-OE, HG7-Scr and HG7-OE cells. (c) BV-2 cells were treated with GCM from GL261, GL261R, GL261-Scr and GL261-OE cells. CD68, CD163, CD204 and CD206 were stained in red, while Arg1, Mrc1, <t>Fizz1</t> and Cd163 were stained in green. Nuclei were stained with DAPI (blue). (d) IHC assays were performed to detect Arg1, Mrc1, Fizz1 and Cd163 in xenograft gliomas formed by GL261-Scr and GL261-OE cells. All images were taken microscopically (20× or 40×). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
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    Photomicrographs of M2 microglia following GCM stimulation of glioma cells in vitro and in vivo. (a) The human microglia HMC3 cells were treated with GCM from LN229, LN229R, HG7 and HG7R cells. (b) HMC3 were treated with GCM from LN229-Scr, LN229-OE, HG7-Scr and HG7-OE cells. (c) BV-2 cells were treated with GCM from GL261, GL261R, GL261-Scr and GL261-OE cells. CD68, CD163, CD204 and CD206 were stained in red, while Arg1, Mrc1, <t>Fizz1</t> and Cd163 were stained in green. Nuclei were stained with DAPI (blue). (d) IHC assays were performed to detect Arg1, Mrc1, Fizz1 and Cd163 in xenograft gliomas formed by GL261-Scr and GL261-OE cells. All images were taken microscopically (20× or 40×). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
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    Photomicrographs of M2 microglia following GCM stimulation of glioma cells in vitro and in vivo. (a) The human microglia HMC3 cells were treated with GCM from LN229, LN229R, HG7 and HG7R cells. (b) HMC3 were treated with GCM from LN229-Scr, LN229-OE, HG7-Scr and HG7-OE cells. (c) BV-2 cells were treated with GCM from GL261, GL261R, GL261-Scr and GL261-OE cells. CD68, CD163, CD204 and CD206 were stained in red, while Arg1, Mrc1, <t>Fizz1</t> and Cd163 were stained in green. Nuclei were stained with DAPI (blue). (d) IHC assays were performed to detect Arg1, Mrc1, Fizz1 and Cd163 in xenograft gliomas formed by GL261-Scr and GL261-OE cells. All images were taken microscopically (20× or 40×). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
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    <t>FIZZ1</t> and YM1 expression in lung epithelial cells. Allergic lung inflammation was induced in RAG2 −/− and γ c −/− mice as mentioned in Fig. 1. FIZZ1 and YM1 expression was analyzed in serial sections of mouse lungs by immunohistochemistry. Photomicrographs (40X magnification) of YM1 (panels a-d) and FIZZ1 (panels e-h) expression in epithelial cells in representative lung sections are shown. (B) The number of YM1+ or FIZZ1+ airways in each group of mice was counted. Data represented as number of airways ± SEM. *p
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    <t>FIZZ1</t> and YM1 expression in lung epithelial cells. Allergic lung inflammation was induced in RAG2 −/− and γ c −/− mice as mentioned in Fig. 1. FIZZ1 and YM1 expression was analyzed in serial sections of mouse lungs by immunohistochemistry. Photomicrographs (40X magnification) of YM1 (panels a-d) and FIZZ1 (panels e-h) expression in epithelial cells in representative lung sections are shown. (B) The number of YM1+ or FIZZ1+ airways in each group of mice was counted. Data represented as number of airways ± SEM. *p
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    <t>Fizz1-</t> and Ym1-expressing macrophages in wound tissue of IL-10 −/− and control mice. Day 5 wounds of control ( A , B ) and IL-10 −/− mice ( C , D ) stained for Fizz1 ( A , C ) or Ym1 ( B , D ); numerous mononuclear cells within the granulation tissue of control and mutant mice stain positive for Fizz1 or Ym1. E: Double labeling for F4/80 (green) and Fizz1 or Ym1 (red) in granulation tissue of IL-10 −/− mice demonstrates Fizz1 and Ym1 expression by macrophages. Scale bars = 100 μm. Original magnifications, ×400 ( E ).
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    <t>Fizz1-</t> and Ym1-expressing macrophages in wound tissue of IL-10 −/− and control mice. Day 5 wounds of control ( A , B ) and IL-10 −/− mice ( C , D ) stained for Fizz1 ( A , C ) or Ym1 ( B , D ); numerous mononuclear cells within the granulation tissue of control and mutant mice stain positive for Fizz1 or Ym1. E: Double labeling for F4/80 (green) and Fizz1 or Ym1 (red) in granulation tissue of IL-10 −/− mice demonstrates Fizz1 and Ym1 expression by macrophages. Scale bars = 100 μm. Original magnifications, ×400 ( E ).
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    <t>Fizz1-</t> and Ym1-expressing macrophages in wound tissue of IL-10 −/− and control mice. Day 5 wounds of control ( A , B ) and IL-10 −/− mice ( C , D ) stained for Fizz1 ( A , C ) or Ym1 ( B , D ); numerous mononuclear cells within the granulation tissue of control and mutant mice stain positive for Fizz1 or Ym1. E: Double labeling for F4/80 (green) and Fizz1 or Ym1 (red) in granulation tissue of IL-10 −/− mice demonstrates Fizz1 and Ym1 expression by macrophages. Scale bars = 100 μm. Original magnifications, ×400 ( E ).
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    Lung macrophages prominently express M2 proteins following IL-4-treatment in intratracheal LPS-exposed WT mice. A : mRNA expression for the M2 markers Arg1 , <t>Fizz1</t> , and Ym1 quantified in the whole lung at days 4 and 6 among intratracheal H 2 O- or LPS-exposed
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    Lung macrophages prominently express M2 proteins following IL-4-treatment in intratracheal LPS-exposed WT mice. A : mRNA expression for the M2 markers Arg1 , <t>Fizz1</t> , and Ym1 quantified in the whole lung at days 4 and 6 among intratracheal H 2 O- or LPS-exposed
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    Image Search Results


    The pleiotropic effects of MTD chemotherapy–treated CAFs mediated through the ELR + chemokine−CXCR-2 paracrine signaling axis. (A) Bar graphs showing the increased endothelial tube formation induced by MTD-doxorubicin– or MTD-paclitaxel–treated BCAF-011 CAFs that could be reduced by treatment with the CXCR-2 inhibitor SB225002 (1 µM) or prevented when LDM-mimetic regimens as described in Fig. 5 (A and C) were used to treat CAFs. Data from two independent experiments ( n = 3 in each group) are shown. (B, left) Representative photomicrographs showing pronounced vascularity in tumors formed by orthotopically implanted BC-011 carcinoma cells along with MTD-doxorubicin (MTD-CAF)– or vehicle (vehicle-CAF)-treated CAFs with or without concurrent intraperitoneal injections of SB225002 (0.5 mg/kg/day) in NOG mice. Also shown are immunohistochemical images of von Willebrand factor (vWF) staining of the endothelial cells in the tumors. Bar, 50 µm. (Right) quantification of microvessel density (MVD) per high-power field (HPF) in tumors. At least 10 different areas were examined in each tissue. Data from one experiment ( n = 3 in each group) are shown. (C, left) The invasive capacities of U937-derived macrophages in response to vehicle-CAFs, MTD-CAFs, or LDM-CAFs in the absence or presence of 1 µM SB225002 in a Transwell invasion assay. Shown are representative immunofluorescence images of the invaded cells, with cell nuclei stained with CYTOX-green. Bars, 100 µm. (Right) The numbers of invaded macrophages. Data from two independent experiments ( n = 3 in each group) are shown. (D, left) Representative immunohistochemical images of F4/80 staining in tumors formed by subcutaneous inoculation of BC-011 cells along with vehicle-, MTD-, or LDM-CAFs in the flank of nude mice. Bar, 50 µm. (Right) Percent F4/80- or Fizz-1–positive cells in the microscopic field. At least 10 different areas were examined in each tissue. Data from two independent experiment ( n = 3 tumor tissues in each group) are shown. (E) BCAF-011 CAFs were treated with vehicle (vehicle-CAF), MTD, (MTD-CAF), or LDM-doxorubicin or paclitaxel (LDM-CAF) as in Fig. 5 and then co-cultivated with BC-011 carcinoma cells in the presence or absence of 1 µM SB225002 in a dual chamber culture apparatus for 5 d, after which the carcinoma cells were subjected to flow cytometric analyses. Shown are the percentages of the CD44 + CD24 −/low carcinoma cells. Data from two independent experiments ( n = 3 in each group) are shown. (F) Freshly sorted CD44 + CD24 −/low BC-011 carcinoma cells were cultured in the presence of human recombinant CXCL-1, -2, -5, and -6 (each at 1 µg/ml) or vehicle in low-attachment culture plates for 10 d, and the diameters of the tumorspheres generated were quantified. Data from two independent experiments ( n = 3 in each group) are shown. (G) The ability of CD44 + CD24 −/low BC-011 carcinoma cells to invade through reconstituted basement membrane in response to human recombinant CXCL-1, -2, -5, and -6 (each at 1 µg/ml) in a Transwell invasion assay. Data from two independent experiments ( n = 3 in each group) are shown. (H) The percentages of CD44 + CD24 −/low cells in BC-011 carcinoma cells co-cultivated with vehicle- or MTD-CAFs along with the neutralizing antibody directed against individual ELR + chemokines. Data from two independent experiments ( n = 3 in each group) are shown. (I) Limiting dilution assay demonstrating the tumorsphere formation efficiency of BC-011 carcinoma cells cultured in the conditioned medium from MTD- or LDM-CAFs. The arrow indicates change of slope of the trend line, suggestive of differential tumorsphere formation ability. Data from one experiment ( n = 6 in each group; mean ± SEM; Student’s t test; ***, P

    Journal: The Journal of Experimental Medicine

    Article Title: Metronomic chemotherapy prevents therapy-induced stromal activation and induction of tumor-initiating cells

    doi: 10.1084/jem.20151665

    Figure Lengend Snippet: The pleiotropic effects of MTD chemotherapy–treated CAFs mediated through the ELR + chemokine−CXCR-2 paracrine signaling axis. (A) Bar graphs showing the increased endothelial tube formation induced by MTD-doxorubicin– or MTD-paclitaxel–treated BCAF-011 CAFs that could be reduced by treatment with the CXCR-2 inhibitor SB225002 (1 µM) or prevented when LDM-mimetic regimens as described in Fig. 5 (A and C) were used to treat CAFs. Data from two independent experiments ( n = 3 in each group) are shown. (B, left) Representative photomicrographs showing pronounced vascularity in tumors formed by orthotopically implanted BC-011 carcinoma cells along with MTD-doxorubicin (MTD-CAF)– or vehicle (vehicle-CAF)-treated CAFs with or without concurrent intraperitoneal injections of SB225002 (0.5 mg/kg/day) in NOG mice. Also shown are immunohistochemical images of von Willebrand factor (vWF) staining of the endothelial cells in the tumors. Bar, 50 µm. (Right) quantification of microvessel density (MVD) per high-power field (HPF) in tumors. At least 10 different areas were examined in each tissue. Data from one experiment ( n = 3 in each group) are shown. (C, left) The invasive capacities of U937-derived macrophages in response to vehicle-CAFs, MTD-CAFs, or LDM-CAFs in the absence or presence of 1 µM SB225002 in a Transwell invasion assay. Shown are representative immunofluorescence images of the invaded cells, with cell nuclei stained with CYTOX-green. Bars, 100 µm. (Right) The numbers of invaded macrophages. Data from two independent experiments ( n = 3 in each group) are shown. (D, left) Representative immunohistochemical images of F4/80 staining in tumors formed by subcutaneous inoculation of BC-011 cells along with vehicle-, MTD-, or LDM-CAFs in the flank of nude mice. Bar, 50 µm. (Right) Percent F4/80- or Fizz-1–positive cells in the microscopic field. At least 10 different areas were examined in each tissue. Data from two independent experiment ( n = 3 tumor tissues in each group) are shown. (E) BCAF-011 CAFs were treated with vehicle (vehicle-CAF), MTD, (MTD-CAF), or LDM-doxorubicin or paclitaxel (LDM-CAF) as in Fig. 5 and then co-cultivated with BC-011 carcinoma cells in the presence or absence of 1 µM SB225002 in a dual chamber culture apparatus for 5 d, after which the carcinoma cells were subjected to flow cytometric analyses. Shown are the percentages of the CD44 + CD24 −/low carcinoma cells. Data from two independent experiments ( n = 3 in each group) are shown. (F) Freshly sorted CD44 + CD24 −/low BC-011 carcinoma cells were cultured in the presence of human recombinant CXCL-1, -2, -5, and -6 (each at 1 µg/ml) or vehicle in low-attachment culture plates for 10 d, and the diameters of the tumorspheres generated were quantified. Data from two independent experiments ( n = 3 in each group) are shown. (G) The ability of CD44 + CD24 −/low BC-011 carcinoma cells to invade through reconstituted basement membrane in response to human recombinant CXCL-1, -2, -5, and -6 (each at 1 µg/ml) in a Transwell invasion assay. Data from two independent experiments ( n = 3 in each group) are shown. (H) The percentages of CD44 + CD24 −/low cells in BC-011 carcinoma cells co-cultivated with vehicle- or MTD-CAFs along with the neutralizing antibody directed against individual ELR + chemokines. Data from two independent experiments ( n = 3 in each group) are shown. (I) Limiting dilution assay demonstrating the tumorsphere formation efficiency of BC-011 carcinoma cells cultured in the conditioned medium from MTD- or LDM-CAFs. The arrow indicates change of slope of the trend line, suggestive of differential tumorsphere formation ability. Data from one experiment ( n = 6 in each group; mean ± SEM; Student’s t test; ***, P

    Article Snippet: Immunohistochemical staining Formalin-fixed, paraffin-embedded human or mouse tumor tissues were processed using standard protocols, stained with anti–α-SMA (for detection of CAFs; α sm-1; Abcam), anti-F4/80 (for detection of TAMs; Hycult Biotech), anti–Fizz-1 (for detection of M2-subtype TAMs; Abcam), or anti–p–STAT-1 (for detection in CAFs; Cell Signaling Technology) and then detected by using a Dako EnVision kit.

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Derivative Assay, Transwell Invasion Assay, Immunofluorescence, Flow Cytometry, Cell Culture, Recombinant, Generated, Limiting Dilution Assay

    Induction of FIZZ1 and YM1. Wild-type mice received DM or MWCNTs (40 μg per mouse) and were sacrificed on day 1, 3, or 7 post-exposure. (A) qRT-PCR. Levels of Fizz1 and Ym1 mRNA from lung tissues were determined, with Gapdh as internal control (mean ± SD, n = 5). (B) Immunoblotting. Levels of FIZZ1 and YM1 proteins in lung tissue lysates were determined ( n = 2). (C) and (D) Time-dependent induction of FIZZ1 (C) and YM1 (D) detected by immunohistochemistry on lung tissue sections. Red indicates positive staining and blue nuclear counterstaining (scale bar: 50 μm). Quantification of positively stained cells is shown as mean ± SD ( n = 4).

    Journal: Nanotoxicology

    Article Title: Macrophage polarization and activation at the interface of multi-walled carbon nanotube-induced pulmonary inflammation and fibrosis

    doi: 10.1080/17435390.2018.1425501

    Figure Lengend Snippet: Induction of FIZZ1 and YM1. Wild-type mice received DM or MWCNTs (40 μg per mouse) and were sacrificed on day 1, 3, or 7 post-exposure. (A) qRT-PCR. Levels of Fizz1 and Ym1 mRNA from lung tissues were determined, with Gapdh as internal control (mean ± SD, n = 5). (B) Immunoblotting. Levels of FIZZ1 and YM1 proteins in lung tissue lysates were determined ( n = 2). (C) and (D) Time-dependent induction of FIZZ1 (C) and YM1 (D) detected by immunohistochemistry on lung tissue sections. Red indicates positive staining and blue nuclear counterstaining (scale bar: 50 μm). Quantification of positively stained cells is shown as mean ± SD ( n = 4).

    Article Snippet: The primary antibodies used were anti-CD86 (Boster, Pleasanton, CA), anti-MHC II (Thermo Fisher Scientific-eBioscience), anti-CD206 (Abcam), anti-CD163 (Abcam), anti-iNOS (Santa Cruz), anti-ARG1 (Santa Cruz), anti-FIZZ1 (Abcam), anti-YM1 (R & D Systems), anti-phospho-STAT1 (Tyr701, Cell Signaling Technology), anti-STAT1 (Cell Signaling Technology), anti-phospho-STAT6 (Y641, Abcam), anti-STAT6 (Cell Signaling Technology), anti-phospho-STAT3 (Tyr705, Cell Signaling Technology), anti-STAT3 (Cell Signaling Technology), anti-IRF5 (Abcam), anti-IRF4 (Proteintech, Rosemont, IL), and anti-Actin (Santa Cruz) antibodies.

    Techniques: Mouse Assay, Quantitative RT-PCR, Immunohistochemistry, Staining

    Rac2 is required for enhanced alternative activation of the macrophage phenotype after bleomycin-induced pulmonary fibrosis. (A B) WT and Rac2-/- mice were given an i.t. challenge with bleomycin and alveolar macrophages were isolated from BALs on day 28 (n = 8–10 mice/group). All the BAL samples/group were pooled to isolate alveolar macrophages and were used for mRNA expression or Western blot or arginase activity. The expression of alternatively activated markers YM1, Fizz1, CD206 and Arginase in alveolar macrophages were analyzed by real-time RT-PCR (A) or Western blot (B). (C) Arginase activity was measured in alveolar macrophages isolated from BALs from bleomycin instilled WT and Rac2-/- mice on day 28 as described in Methods. Graphs in A and C represent mean ± SEM with n = 4. One-way ANOVA with post-hoc Tukey’s multiple comparison tests, **p ≤ 0.01 and ***p ≤ 0.001 when WT bleomycin treated group was compared to saline treated groups or the Rac2-/- bleomycin treated group. Experiment was repeated twice with similar results.

    Journal: PLoS ONE

    Article Title: Rac2 is required for alternative macrophage activation and bleomycin induced pulmonary fibrosis; a macrophage autonomous phenotype

    doi: 10.1371/journal.pone.0182851

    Figure Lengend Snippet: Rac2 is required for enhanced alternative activation of the macrophage phenotype after bleomycin-induced pulmonary fibrosis. (A B) WT and Rac2-/- mice were given an i.t. challenge with bleomycin and alveolar macrophages were isolated from BALs on day 28 (n = 8–10 mice/group). All the BAL samples/group were pooled to isolate alveolar macrophages and were used for mRNA expression or Western blot or arginase activity. The expression of alternatively activated markers YM1, Fizz1, CD206 and Arginase in alveolar macrophages were analyzed by real-time RT-PCR (A) or Western blot (B). (C) Arginase activity was measured in alveolar macrophages isolated from BALs from bleomycin instilled WT and Rac2-/- mice on day 28 as described in Methods. Graphs in A and C represent mean ± SEM with n = 4. One-way ANOVA with post-hoc Tukey’s multiple comparison tests, **p ≤ 0.01 and ***p ≤ 0.001 when WT bleomycin treated group was compared to saline treated groups or the Rac2-/- bleomycin treated group. Experiment was repeated twice with similar results.

    Article Snippet: Membranes were incubated with antibodies against FIZZ1 (Abcam), Arg (BD Biosciences) and CD206 (Abcam) antibodies.

    Techniques: Activation Assay, Mouse Assay, Isolation, Expressing, Western Blot, Activity Assay, Quantitative RT-PCR

    α 4 β 1 integrin is required for bleomycin-induced pulmonary fibrosis. (A) WT and α4Y991A mice were given an i.t. challenge with bleomycin (n = 8 mice / group), and whole lungs were isolated on day 28 and amount of collagen was assessed by hydroxyproline assay (n = 4 mice/group) (A), Sirius red staining of histological sections, Original magnification 20X (B), mRNA expression of fibrosis related genes (C) in saline and bleomycin instilled lungs isolated from WT and α4Y991A mice (n = 3 samples/group). For mRNA analysis and Western blot, BAL samples from n = 8 mice / group were pooled together to isolate alveolar macrophages for either mRNA expression or Western blot. (D) Expression of alternatively activated markers YM1, Fizz1, CD206 and Arginase in alveolar macrophages isolated from BALs from bleomycin instilled WT and α4Y991A mice on day 28 and analyzed by real-time RT-PCR. (E) Western blot analysis of M2 marker genes in bleomycin challenged WT and α4Y991A mice. Graphs represent mean ± SEM with n = 4 samples for A, n = 3 for C and n = 4 for D. One-way ANOVA with post-hoc Tukey’s multiple comparison tests, *p ≤ 0.05, ***p ≤ 0.001 when WT bleomycin treated group was compared to saline treated groups or the Rac2-/- bleomycin treated group. Experiment was repeated twice with same results.

    Journal: PLoS ONE

    Article Title: Rac2 is required for alternative macrophage activation and bleomycin induced pulmonary fibrosis; a macrophage autonomous phenotype

    doi: 10.1371/journal.pone.0182851

    Figure Lengend Snippet: α 4 β 1 integrin is required for bleomycin-induced pulmonary fibrosis. (A) WT and α4Y991A mice were given an i.t. challenge with bleomycin (n = 8 mice / group), and whole lungs were isolated on day 28 and amount of collagen was assessed by hydroxyproline assay (n = 4 mice/group) (A), Sirius red staining of histological sections, Original magnification 20X (B), mRNA expression of fibrosis related genes (C) in saline and bleomycin instilled lungs isolated from WT and α4Y991A mice (n = 3 samples/group). For mRNA analysis and Western blot, BAL samples from n = 8 mice / group were pooled together to isolate alveolar macrophages for either mRNA expression or Western blot. (D) Expression of alternatively activated markers YM1, Fizz1, CD206 and Arginase in alveolar macrophages isolated from BALs from bleomycin instilled WT and α4Y991A mice on day 28 and analyzed by real-time RT-PCR. (E) Western blot analysis of M2 marker genes in bleomycin challenged WT and α4Y991A mice. Graphs represent mean ± SEM with n = 4 samples for A, n = 3 for C and n = 4 for D. One-way ANOVA with post-hoc Tukey’s multiple comparison tests, *p ≤ 0.05, ***p ≤ 0.001 when WT bleomycin treated group was compared to saline treated groups or the Rac2-/- bleomycin treated group. Experiment was repeated twice with same results.

    Article Snippet: Membranes were incubated with antibodies against FIZZ1 (Abcam), Arg (BD Biosciences) and CD206 (Abcam) antibodies.

    Techniques: Mouse Assay, Isolation, Hydroxyproline Assay, Staining, Expressing, Western Blot, Quantitative RT-PCR, Marker

    DHT enhances M2 gene expression, YM1 and FIZZ1 production in BMM. Bone marrow from female and male C57BL/6 mice was cultured for 10 days with M-CSF (40 ng/mL) to obtain BMM. (A) Validation of the specificity of anti-AR antibody. AM from female and male mice were cultured for 24 h in the presence or absence of the AR-degrading drug, ASC-J9 (10 μM), left panel. AR expression in BMM from C57BL/6 female and male mice, middle panel. Comparison of AR expression in BMM and AM, right panel. (B) BMM were cultured for 10 days with 40 ng/mL of GM-CSF or M-CSF in 6-well plates, cells were collected and viability determined. F4/80 + cells were selected in the live population. Dot plots of CD11c + Siglec F + cells from F4/80 + cells cultured with GM-CSF or M-CSF. (C) Analysis of double-positive CD11c + Siglec F + macrophages with GM-CSF or M-CSF (D) Analysis of AR + cells from F4/80 + cells cultured with GM-CSF or M-CSF. (E) BMM were pretreated overnight with the indicated concentrations of DHT and then stimulated with IL-4 (1 ng/mL) for 48 h. Expression of the indicated M2 genes was determined by qPCR using the 2 -ΔΔCT method, and compared to the amount of mRNA in the male IL-4 sample (= 100%). (F) Expression of YM1 and FIZZ1 protein was determined by Western blot in the same BMM supernatants after 48 h of IL-4 stimulation. Equal volumes of supernatant were loaded in each lane. Densitometry of the specific bands was normalized to the amount of protein in male IL-4 (= 100%). Results are representative of four independent in vitro experiments. † p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Androgen and androgen receptor as enhancers of M2 macrophage polarization in allergic lung inflammation

    doi: 10.4049/jimmunol.1800352

    Figure Lengend Snippet: DHT enhances M2 gene expression, YM1 and FIZZ1 production in BMM. Bone marrow from female and male C57BL/6 mice was cultured for 10 days with M-CSF (40 ng/mL) to obtain BMM. (A) Validation of the specificity of anti-AR antibody. AM from female and male mice were cultured for 24 h in the presence or absence of the AR-degrading drug, ASC-J9 (10 μM), left panel. AR expression in BMM from C57BL/6 female and male mice, middle panel. Comparison of AR expression in BMM and AM, right panel. (B) BMM were cultured for 10 days with 40 ng/mL of GM-CSF or M-CSF in 6-well plates, cells were collected and viability determined. F4/80 + cells were selected in the live population. Dot plots of CD11c + Siglec F + cells from F4/80 + cells cultured with GM-CSF or M-CSF. (C) Analysis of double-positive CD11c + Siglec F + macrophages with GM-CSF or M-CSF (D) Analysis of AR + cells from F4/80 + cells cultured with GM-CSF or M-CSF. (E) BMM were pretreated overnight with the indicated concentrations of DHT and then stimulated with IL-4 (1 ng/mL) for 48 h. Expression of the indicated M2 genes was determined by qPCR using the 2 -ΔΔCT method, and compared to the amount of mRNA in the male IL-4 sample (= 100%). (F) Expression of YM1 and FIZZ1 protein was determined by Western blot in the same BMM supernatants after 48 h of IL-4 stimulation. Equal volumes of supernatant were loaded in each lane. Densitometry of the specific bands was normalized to the amount of protein in male IL-4 (= 100%). Results are representative of four independent in vitro experiments. † p

    Article Snippet: Proteins were transferred onto PVDF membranes (Bio-Rad, Hercules, CA), and probed with antibodies against YM1 (STEMCELL Technologies) and FIZZ1 (Abcam, Cambridge, MA).

    Techniques: Expressing, Mouse Assay, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, In Vitro

    FIZZ1 induces myofibroblast differentiation. A: Fully differentiated 3T3-L1 adipocytes were treated with 25 ng/mL FIZZ1 for 7 days. The indicated genes were analyzed by RT-qPCR. B: α-SMA protein induction with indicated doses of FIZZ1 was analyzed

    Journal: The American Journal of Pathology

    Article Title: FIZZ1-Induced Myofibroblast Transdifferentiation from Adipocytes and Its Potential Role in Dermal Fibrosis and Lipoatrophy

    doi: 10.1016/j.ajpath.2015.06.005

    Figure Lengend Snippet: FIZZ1 induces myofibroblast differentiation. A: Fully differentiated 3T3-L1 adipocytes were treated with 25 ng/mL FIZZ1 for 7 days. The indicated genes were analyzed by RT-qPCR. B: α-SMA protein induction with indicated doses of FIZZ1 was analyzed

    Article Snippet: The following antibodies were used: anti–α-SMA (Sigma-Aldrich), anti-collagen type I (Biodesign International, Saco, ME), anti–PPAR-γ (Cell Signaling Technology, Inc., Danvers, MA), anti-FABP4 (Cayman, Ann Arbor, MI), anti-FIZZ1 (R & D Systems, Minneapolis, MN), anti-Notch1 (Cell Signaling Technology, Inc.), and horseradish peroxidase-conjugated anti–glyceraldehyde-3-phosphate dehydrogenase (Sigma-Aldrich).

    Techniques: Quantitative RT-PCR

    FIZZ1 expression and localization in BLM-induced dermal fibrosis model. A: Twenty-one days after daily BLM subcutaneous injections, skin tissue RNA was analyzed for α-SMA and type I procollagen by RT-qPCR for fibrogenic gene expression. B: FIZZ1

    Journal: The American Journal of Pathology

    Article Title: FIZZ1-Induced Myofibroblast Transdifferentiation from Adipocytes and Its Potential Role in Dermal Fibrosis and Lipoatrophy

    doi: 10.1016/j.ajpath.2015.06.005

    Figure Lengend Snippet: FIZZ1 expression and localization in BLM-induced dermal fibrosis model. A: Twenty-one days after daily BLM subcutaneous injections, skin tissue RNA was analyzed for α-SMA and type I procollagen by RT-qPCR for fibrogenic gene expression. B: FIZZ1

    Article Snippet: The following antibodies were used: anti–α-SMA (Sigma-Aldrich), anti-collagen type I (Biodesign International, Saco, ME), anti–PPAR-γ (Cell Signaling Technology, Inc., Danvers, MA), anti-FABP4 (Cayman, Ann Arbor, MI), anti-FIZZ1 (R & D Systems, Minneapolis, MN), anti-Notch1 (Cell Signaling Technology, Inc.), and horseradish peroxidase-conjugated anti–glyceraldehyde-3-phosphate dehydrogenase (Sigma-Aldrich).

    Techniques: Expressing, Quantitative RT-PCR

    Notch1 mediates FIZZ1 effect on adipocyte transdifferentiation. Fully differentiated adipocytes were treated with 25 ng/mL FIZZ1 for 24 hours. A: Notch1 mRNA was analyzed by RT-qPCR. The adipocytes were transfected with 100 nmol/L Notch1 or control siRNA

    Journal: The American Journal of Pathology

    Article Title: FIZZ1-Induced Myofibroblast Transdifferentiation from Adipocytes and Its Potential Role in Dermal Fibrosis and Lipoatrophy

    doi: 10.1016/j.ajpath.2015.06.005

    Figure Lengend Snippet: Notch1 mediates FIZZ1 effect on adipocyte transdifferentiation. Fully differentiated adipocytes were treated with 25 ng/mL FIZZ1 for 24 hours. A: Notch1 mRNA was analyzed by RT-qPCR. The adipocytes were transfected with 100 nmol/L Notch1 or control siRNA

    Article Snippet: The following antibodies were used: anti–α-SMA (Sigma-Aldrich), anti-collagen type I (Biodesign International, Saco, ME), anti–PPAR-γ (Cell Signaling Technology, Inc., Danvers, MA), anti-FABP4 (Cayman, Ann Arbor, MI), anti-FIZZ1 (R & D Systems, Minneapolis, MN), anti-Notch1 (Cell Signaling Technology, Inc.), and horseradish peroxidase-conjugated anti–glyceraldehyde-3-phosphate dehydrogenase (Sigma-Aldrich).

    Techniques: Quantitative RT-PCR, Transfection

    FIZZ1 induction and induced adipocyte dedifferentiation. A: 3T3-L1 preadipocytes were grown to confluence in SM and then induced for adipocyte differentiation for 6 days without (DM) or with subsequent FIZZ1 treatment (DM + FIZZ1) for

    Journal: The American Journal of Pathology

    Article Title: FIZZ1-Induced Myofibroblast Transdifferentiation from Adipocytes and Its Potential Role in Dermal Fibrosis and Lipoatrophy

    doi: 10.1016/j.ajpath.2015.06.005

    Figure Lengend Snippet: FIZZ1 induction and induced adipocyte dedifferentiation. A: 3T3-L1 preadipocytes were grown to confluence in SM and then induced for adipocyte differentiation for 6 days without (DM) or with subsequent FIZZ1 treatment (DM + FIZZ1) for

    Article Snippet: The following antibodies were used: anti–α-SMA (Sigma-Aldrich), anti-collagen type I (Biodesign International, Saco, ME), anti–PPAR-γ (Cell Signaling Technology, Inc., Danvers, MA), anti-FABP4 (Cayman, Ann Arbor, MI), anti-FIZZ1 (R & D Systems, Minneapolis, MN), anti-Notch1 (Cell Signaling Technology, Inc.), and horseradish peroxidase-conjugated anti–glyceraldehyde-3-phosphate dehydrogenase (Sigma-Aldrich).

    Techniques:

    FIZZ1 inhibits adipocyte gene expression. Fully differentiated 3T3-L1 adipocytes were treated with 25 ng/mL FIZZ1 for 3 days. The indicated adipocyte specific markers were analyzed by RT-qPCR. The resulting mRNA levels were expressed as 2 −ΔΔC

    Journal: The American Journal of Pathology

    Article Title: FIZZ1-Induced Myofibroblast Transdifferentiation from Adipocytes and Its Potential Role in Dermal Fibrosis and Lipoatrophy

    doi: 10.1016/j.ajpath.2015.06.005

    Figure Lengend Snippet: FIZZ1 inhibits adipocyte gene expression. Fully differentiated 3T3-L1 adipocytes were treated with 25 ng/mL FIZZ1 for 3 days. The indicated adipocyte specific markers were analyzed by RT-qPCR. The resulting mRNA levels were expressed as 2 −ΔΔC

    Article Snippet: The following antibodies were used: anti–α-SMA (Sigma-Aldrich), anti-collagen type I (Biodesign International, Saco, ME), anti–PPAR-γ (Cell Signaling Technology, Inc., Danvers, MA), anti-FABP4 (Cayman, Ann Arbor, MI), anti-FIZZ1 (R & D Systems, Minneapolis, MN), anti-Notch1 (Cell Signaling Technology, Inc.), and horseradish peroxidase-conjugated anti–glyceraldehyde-3-phosphate dehydrogenase (Sigma-Aldrich).

    Techniques: Expressing, Quantitative RT-PCR

    Histological examination of lung tissue. Tissues from pre-terminal bronchioles were analyzed for morphologic features, mucosubstances in mucous cells and the presence of some proteins associated with allergic lung inflammation. (A-C) H E staining. (D-E) AB/PAS staining. Arrows depict AB/PAS-stained mucosubstances in airway epithelium. (G-I) Immunohistological staining using anti Ym1/Ym2 antibodies. Arrows depict stained Ym1/Ym2. (J-L) Immunohistological staining using anti FIZZ1 antibodies. Arrows depict FIZZ in the epithelium. Abbreviations: AB/PAS, Alcian blue-Periodic acid schiff double stain; AL, airway lumen; ap, alveolar parenchyma; bv, blood vessel; e, airway surface epithelium; H E, hematoxylin and eosin stain.

    Journal: Proteomics

    Article Title: Adjuvant Effects of Ambient Particulate Matter Monitored by Proteomics of Bronchoalveolar Lavage Fluid

    doi: 10.1002/pmic.200900573

    Figure Lengend Snippet: Histological examination of lung tissue. Tissues from pre-terminal bronchioles were analyzed for morphologic features, mucosubstances in mucous cells and the presence of some proteins associated with allergic lung inflammation. (A-C) H E staining. (D-E) AB/PAS staining. Arrows depict AB/PAS-stained mucosubstances in airway epithelium. (G-I) Immunohistological staining using anti Ym1/Ym2 antibodies. Arrows depict stained Ym1/Ym2. (J-L) Immunohistological staining using anti FIZZ1 antibodies. Arrows depict FIZZ in the epithelium. Abbreviations: AB/PAS, Alcian blue-Periodic acid schiff double stain; AL, airway lumen; ap, alveolar parenchyma; bv, blood vessel; e, airway surface epithelium; H E, hematoxylin and eosin stain.

    Article Snippet: The mucosa changes were accompanied by immunohistological evidence of over expression of Ym1/2 and FIZZ1 ( , Ym1/2, FIZZ staining).

    Techniques: Staining, H&E Stain

    Lung macrophages prominently express M2 proteins following IL-4-treatment in intratracheal LPS-exposed WT mice. A : mRNA expression for the M2 markers Arg1 , Fizz1 , and Ym1 quantified in the whole lung at days 4 and 6 among intratracheal H 2 O- or LPS-exposed

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Enhanced resolution of experimental ARDS through IL-4-mediated lung macrophage reprogramming

    doi: 10.1152/ajplung.00419.2015

    Figure Lengend Snippet: Lung macrophages prominently express M2 proteins following IL-4-treatment in intratracheal LPS-exposed WT mice. A : mRNA expression for the M2 markers Arg1 , Fizz1 , and Ym1 quantified in the whole lung at days 4 and 6 among intratracheal H 2 O- or LPS-exposed

    Article Snippet: Then, 1 × 106 surface-stained cells were washed, fixed, and permeabilized with a Cytofix/Cytoperm kit (BD Biosciences) before intracellular staining for 30 min with rabbit anti-mouse primary antibodies to FIZZ1 (1:100; Peprotech).

    Techniques: Mouse Assay, Expressing

    The effect of ECM and mechanical stimulation upon the myoblast secretome. C2C12 cells were cultured in proliferation media or allowed to form myotubes in differentiation media culture. Myoblasts or myotubes were treated with 200 μg/mL of ECM degradation products for 18 h, after which the media were replaced with serum-free, ECM-free media and the cells were subjected to mechanical strain. Conditioned media were collected and added to bone marrow-derived macrophage for 18 h and the cells were fixed for immunolabeling. (A) Exposure times were normalized to cytokine stimulated controls. (B, D) The secretome of proliferating myoblasts promotes an iNOS−/Fizz1+ macrophage phenotype, (C) however, differentiated myotubes do not promote the same effect. Treating myotubes with ECM degradation products, however, alters their secretome allowing them to promote a Fizz1+ macrophage phenotype. (E) This response is augmented when ECM-treated myotubes are subjected to mechanical strain. (F) Percentage of iNOS- and Fizz1-positive macrophages were quantified using CellProfiler. (# p

    Journal: Tissue Engineering. Part A

    Article Title: The Effect of Mechanical Loading Upon Extracellular Matrix Bioscaffold-Mediated Skeletal Muscle Remodeling

    doi: 10.1089/ten.tea.2017.0011

    Figure Lengend Snippet: The effect of ECM and mechanical stimulation upon the myoblast secretome. C2C12 cells were cultured in proliferation media or allowed to form myotubes in differentiation media culture. Myoblasts or myotubes were treated with 200 μg/mL of ECM degradation products for 18 h, after which the media were replaced with serum-free, ECM-free media and the cells were subjected to mechanical strain. Conditioned media were collected and added to bone marrow-derived macrophage for 18 h and the cells were fixed for immunolabeling. (A) Exposure times were normalized to cytokine stimulated controls. (B, D) The secretome of proliferating myoblasts promotes an iNOS−/Fizz1+ macrophage phenotype, (C) however, differentiated myotubes do not promote the same effect. Treating myotubes with ECM degradation products, however, alters their secretome allowing them to promote a Fizz1+ macrophage phenotype. (E) This response is augmented when ECM-treated myotubes are subjected to mechanical strain. (F) Percentage of iNOS- and Fizz1-positive macrophages were quantified using CellProfiler. (# p

    Article Snippet: After the blocking step, tissue sections were incubated with the following primary antibodies diluted in the blocking solution for 16 h at 4°C: (1) anti-iNOS (abcam) and (2) anti-Fizz1 (Peprotech).

    Techniques: Cell Culture, Derivative Assay, Immunolabeling

    Cyclic mechanical strain of bone marrow-derived macrophages. Macrophages were subjected to 10% mechanical strain for 5 h using the FlexCell system. Five-hour mechanical strain resulted in an F480+/iNOS−/Fizz1+ macrophage phenotype, suggesting that mechanical strain alone can promote macrophage activation toward a proremodeling phenotype. (* p

    Journal: Tissue Engineering. Part A

    Article Title: The Effect of Mechanical Loading Upon Extracellular Matrix Bioscaffold-Mediated Skeletal Muscle Remodeling

    doi: 10.1089/ten.tea.2017.0011

    Figure Lengend Snippet: Cyclic mechanical strain of bone marrow-derived macrophages. Macrophages were subjected to 10% mechanical strain for 5 h using the FlexCell system. Five-hour mechanical strain resulted in an F480+/iNOS−/Fizz1+ macrophage phenotype, suggesting that mechanical strain alone can promote macrophage activation toward a proremodeling phenotype. (* p

    Article Snippet: After the blocking step, tissue sections were incubated with the following primary antibodies diluted in the blocking solution for 16 h at 4°C: (1) anti-iNOS (abcam) and (2) anti-Fizz1 (Peprotech).

    Techniques: Derivative Assay, Activation Assay

    Marophage and myoblast response to hind limb unloading following ECM-mediated VML remodeling. (A) Hind limb unloading results in an increased infiltration of iNOS+ macrophages at 14 days following implantation and (B) decreased Fizz1+ macrophage infiltration in contrast to the (C, D) normal ambulation control. (E) Quantification of the Fizz1+:iNOS+ macrophage ratio across all animals showed a significant decrease following 14 days of hind limb unloading (** p

    Journal: Tissue Engineering. Part A

    Article Title: The Effect of Mechanical Loading Upon Extracellular Matrix Bioscaffold-Mediated Skeletal Muscle Remodeling

    doi: 10.1089/ten.tea.2017.0011

    Figure Lengend Snippet: Marophage and myoblast response to hind limb unloading following ECM-mediated VML remodeling. (A) Hind limb unloading results in an increased infiltration of iNOS+ macrophages at 14 days following implantation and (B) decreased Fizz1+ macrophage infiltration in contrast to the (C, D) normal ambulation control. (E) Quantification of the Fizz1+:iNOS+ macrophage ratio across all animals showed a significant decrease following 14 days of hind limb unloading (** p

    Article Snippet: After the blocking step, tissue sections were incubated with the following primary antibodies diluted in the blocking solution for 16 h at 4°C: (1) anti-iNOS (abcam) and (2) anti-Fizz1 (Peprotech).

    Techniques:

    STAT6 signaling pathway involvement in gefitinib-mediated inhibition of M2-like polarization of macrophages. (A) RAW 264.7 cells were treated with IL-13 (10 ng/mL) with or without gefitinib for the indicated times. Western blots of STAT6 and activated p-STAT6 in cell lysates. (B) Western blot was performed to measure the protein level of Mrc1, Arg1, Ym1, and Fizz1. BMDMs were stimulated with IL-13 (10 ng/mL) and treated with or without gefitinib for 24 h. Actin was used as a loading control. The results are presented as the mean±SEM of triplicate determinations. * P

    Journal: Acta Pharmacologica Sinica

    Article Title: Gefitinib inhibits M2-like polarization of tumor-associated macrophages in Lewis lung cancer by targeting the STAT6 signaling pathway

    doi: 10.1038/aps.2017.124

    Figure Lengend Snippet: STAT6 signaling pathway involvement in gefitinib-mediated inhibition of M2-like polarization of macrophages. (A) RAW 264.7 cells were treated with IL-13 (10 ng/mL) with or without gefitinib for the indicated times. Western blots of STAT6 and activated p-STAT6 in cell lysates. (B) Western blot was performed to measure the protein level of Mrc1, Arg1, Ym1, and Fizz1. BMDMs were stimulated with IL-13 (10 ng/mL) and treated with or without gefitinib for 24 h. Actin was used as a loading control. The results are presented as the mean±SEM of triplicate determinations. * P

    Article Snippet: Antibodies against Mrc1, Arg1, Fizz1 and actin were purchased from Santa Cruz Biotechnology (CA, USA).

    Techniques: Inhibition, Western Blot

    Gefitinib prevents Lewis lung cancer formation in vivo by targeting macrophages. C57BL/6 mice were injected intravenously and were treated with gefitinib (75 mg/kg). After 21 d, animals were euthanized and the lungs were analyzed. (A) The average body weight of each group is expressed as the mean±SD ( n =6). (B) Quantitative analysis of lung metastasis nodules ( n =6). (C) The percentage of M2 surface markers F4/80 + CD206 + was analyzed by FACS analysis. (D) Quantitative real-time PCR was performed to assess the mRNA levels of the M2-marker genes Mrc1, Arg1, and Fizz1. The results are presented as the mean±SEM * P

    Journal: Acta Pharmacologica Sinica

    Article Title: Gefitinib inhibits M2-like polarization of tumor-associated macrophages in Lewis lung cancer by targeting the STAT6 signaling pathway

    doi: 10.1038/aps.2017.124

    Figure Lengend Snippet: Gefitinib prevents Lewis lung cancer formation in vivo by targeting macrophages. C57BL/6 mice were injected intravenously and were treated with gefitinib (75 mg/kg). After 21 d, animals were euthanized and the lungs were analyzed. (A) The average body weight of each group is expressed as the mean±SD ( n =6). (B) Quantitative analysis of lung metastasis nodules ( n =6). (C) The percentage of M2 surface markers F4/80 + CD206 + was analyzed by FACS analysis. (D) Quantitative real-time PCR was performed to assess the mRNA levels of the M2-marker genes Mrc1, Arg1, and Fizz1. The results are presented as the mean±SEM * P

    Article Snippet: Antibodies against Mrc1, Arg1, Fizz1 and actin were purchased from Santa Cruz Biotechnology (CA, USA).

    Techniques: In Vivo, Mouse Assay, Injection, FACS, Real-time Polymerase Chain Reaction, Marker

    Switch to M2a polarization in IKKβ cKO mice at mid-infection stage. M2a activation occurs in IKKβ cKO mice after sublethal i.d. infection with 10 6 Ft . LVS as evidenced by expression of ( A ) CD206 ( B ) IL-10 ( C ) FIZZ1/Relmα and ( D ) Arg-1 in a flow cytometry time course experiment. Statistical analysis was performed by one-way ANOVA followed by Tukey’s ad hoc post-test. (n for IKK f/f , IKKα cKO, IKKβ cKO on day 0∶5/5/5; day 2∶4/4/4; day 6∶4/4/4; day 8∶4/4/3). Results are representative of at least two independent experiments. Bars represent the mean ± SD. **P

    Journal: PLoS ONE

    Article Title: IKK? in Myeloid Cells Controls the Host Response to Lethal and Sublethal Francisella tularensis LVS Infection

    doi: 10.1371/journal.pone.0054124

    Figure Lengend Snippet: Switch to M2a polarization in IKKβ cKO mice at mid-infection stage. M2a activation occurs in IKKβ cKO mice after sublethal i.d. infection with 10 6 Ft . LVS as evidenced by expression of ( A ) CD206 ( B ) IL-10 ( C ) FIZZ1/Relmα and ( D ) Arg-1 in a flow cytometry time course experiment. Statistical analysis was performed by one-way ANOVA followed by Tukey’s ad hoc post-test. (n for IKK f/f , IKKα cKO, IKKβ cKO on day 0∶5/5/5; day 2∶4/4/4; day 6∶4/4/4; day 8∶4/4/3). Results are representative of at least two independent experiments. Bars represent the mean ± SD. **P

    Article Snippet: LVS (Rabbit polyclonal, a generous gift of Dr. Jorge Benach), APC-IFN-γ (XMG1.2, BD Biosciences), PE-IL-10 (JES5-16E3, BD Biosciences), APC-IL-12p40/70 (C15.6, BD Biosciences), FITC-pan-Neutrophils/Ly-6B.2 (7/4, AbD Serotec, Raleigh, NC), PerCP-Cy5.5-NK1.1 (PK136, BD Biosciences), RELMα/FIZZ1 (Rabbit Polyclonal, Abcam, Cambridge, MA), PE-Donkey anti rabbit IgG (Abcam), PE-Donkey anti goat IgG (Abcam).

    Techniques: Mouse Assay, Infection, Activation Assay, Expressing, Flow Cytometry, Cytometry

    Moderate dose of XBJ (5 mg/mL) enhanced the expression of M2 phenotype markers of mouse peritoneal macrophages. Peritoneal macrophages of mice were, respectively, treated with 0 mg/mL, 1 mg/mL, 5 mg/mL, 10 mg/mL, 50 mg/mL, and 100 mg/mL XBJ in the presence of 100 ng/mL LPS for 6 h, 24 h, or 48 h. Cytokines levels including TNF- α , IL-6, and IL-10 in the medium were determined by ELISA (a). F4/80, CD11c, and CD206 expressions were determined by flow cytometry. The group of F4/80 + CD11c + CD206 − was classified as M1 and F4/80 + CD11c − CD206 + as M2 (b). The treated macrophages were lysed and levels of Arg1, Ym1, and Fizz1 were measured by western blot (c). Data represented the mean ± SEM of independent experiment in triplicate. ∗ p

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Xuebijing Injection Promotes M2 Polarization of Macrophages and Improves Survival Rate in Septic Mice

    doi: 10.1155/2015/352642

    Figure Lengend Snippet: Moderate dose of XBJ (5 mg/mL) enhanced the expression of M2 phenotype markers of mouse peritoneal macrophages. Peritoneal macrophages of mice were, respectively, treated with 0 mg/mL, 1 mg/mL, 5 mg/mL, 10 mg/mL, 50 mg/mL, and 100 mg/mL XBJ in the presence of 100 ng/mL LPS for 6 h, 24 h, or 48 h. Cytokines levels including TNF- α , IL-6, and IL-10 in the medium were determined by ELISA (a). F4/80, CD11c, and CD206 expressions were determined by flow cytometry. The group of F4/80 + CD11c + CD206 − was classified as M1 and F4/80 + CD11c − CD206 + as M2 (b). The treated macrophages were lysed and levels of Arg1, Ym1, and Fizz1 were measured by western blot (c). Data represented the mean ± SEM of independent experiment in triplicate. ∗ p

    Article Snippet: Mouse anti-Arg1 was purchased from BD Biosciences (San Jose, CA), mouse Fizz1 antibody was purchased from R & D Systems (Minneapolis, Minn), anti-Ym1 was purchased from Stemcell Technologies (Vancouver, Canada), and anti-β -actin was purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Expressing, Mouse Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cytometry, Western Blot

    The lung HYP content was measured in GFP-FIZZ1 chimera lung samples after BLM or PBS treatment. Data were shown as percentages of their respective PBS controls. The mean values were shown from 2 mice each group. *P

    Journal: PLoS ONE

    Article Title: The In Vivo Fibrotic Role of FIZZ1 in Pulmonary Fibrosis

    doi: 10.1371/journal.pone.0088362

    Figure Lengend Snippet: The lung HYP content was measured in GFP-FIZZ1 chimera lung samples after BLM or PBS treatment. Data were shown as percentages of their respective PBS controls. The mean values were shown from 2 mice each group. *P

    Article Snippet: The following primary antibodies were used: anti-rat FIZZ1 (Alexis Biochemicals, San Diego, CA), anti-α-SMA (Sigma), anti-rat or mouse type I collagen (Biodesign International, Saco, ME), and HRP-conjugated anti GAPDH (Abcam, Cambridge, MA).

    Techniques: Mouse Assay

    FIZZ1 effects on BM cell recruitment. (A) Fresh isolated whole BM cells from day 7 BLM (“BLM-BM”) or PBS (“CON-BM”) treated mice were preloaded with fluorescent dye, and then plated into 5 µm-inserts in 24-well transwell plates. After 2 hours of incubation with the indicated doses of FIZZ1 in the lower chambers, the cells that have migrated to the lower chambers were quantitated by measuring the fluorescence with an excitation and emission wavelengths of 494 and 517 nm, respectively. The results were normalized to the controls and expressed as percentages of controls, and shown as mean ± SE (n = 3). In (B) purified BM-derived DC from day 7 BLM (“BLM-BMDC”) or PBS (“CON-BMDC”) treated mice were similarly analyzed as in (A) for migration to media only (“None”) or to 50 ng/ml FIZZ1 in the lower chambers. The results were expressed as in (A). (C) BM from GFP transgenic mice were transplanted into WT or FIZZ1 KO mice to create GFP BM chimera mice of the respective recipient strain. After stable engraftment the mice were treated with BLM and 7 days later were analyzed for GFP expression in the lung cell population by flow cytometry. Three populations were discerned corresponding to the cells with undetectable (R1), low (R2) or high (R3) GFP fluorescence. One representative data was shown from two individual experiments, and the combined lung cells from two mice were used for flow cytometry in each group. Circulating FIZZ1 protein was measured in the sera (1∶50 dilution) by ELISA assay 21 days after BLM or PBS treatment (D). Results were shown as mean ± SE with 5–6 mice in each group. (E) Effect of FIZZ1 on migration of primary cultured mouse lung fibroblasts was also analyzed as described in legend for (A). *P

    Journal: PLoS ONE

    Article Title: The In Vivo Fibrotic Role of FIZZ1 in Pulmonary Fibrosis

    doi: 10.1371/journal.pone.0088362

    Figure Lengend Snippet: FIZZ1 effects on BM cell recruitment. (A) Fresh isolated whole BM cells from day 7 BLM (“BLM-BM”) or PBS (“CON-BM”) treated mice were preloaded with fluorescent dye, and then plated into 5 µm-inserts in 24-well transwell plates. After 2 hours of incubation with the indicated doses of FIZZ1 in the lower chambers, the cells that have migrated to the lower chambers were quantitated by measuring the fluorescence with an excitation and emission wavelengths of 494 and 517 nm, respectively. The results were normalized to the controls and expressed as percentages of controls, and shown as mean ± SE (n = 3). In (B) purified BM-derived DC from day 7 BLM (“BLM-BMDC”) or PBS (“CON-BMDC”) treated mice were similarly analyzed as in (A) for migration to media only (“None”) or to 50 ng/ml FIZZ1 in the lower chambers. The results were expressed as in (A). (C) BM from GFP transgenic mice were transplanted into WT or FIZZ1 KO mice to create GFP BM chimera mice of the respective recipient strain. After stable engraftment the mice were treated with BLM and 7 days later were analyzed for GFP expression in the lung cell population by flow cytometry. Three populations were discerned corresponding to the cells with undetectable (R1), low (R2) or high (R3) GFP fluorescence. One representative data was shown from two individual experiments, and the combined lung cells from two mice were used for flow cytometry in each group. Circulating FIZZ1 protein was measured in the sera (1∶50 dilution) by ELISA assay 21 days after BLM or PBS treatment (D). Results were shown as mean ± SE with 5–6 mice in each group. (E) Effect of FIZZ1 on migration of primary cultured mouse lung fibroblasts was also analyzed as described in legend for (A). *P

    Article Snippet: The following primary antibodies were used: anti-rat FIZZ1 (Alexis Biochemicals, San Diego, CA), anti-α-SMA (Sigma), anti-rat or mouse type I collagen (Biodesign International, Saco, ME), and HRP-conjugated anti GAPDH (Abcam, Cambridge, MA).

    Techniques: Isolation, Mouse Assay, Incubation, Fluorescence, Purification, Derivative Assay, Migration, Transgenic Assay, Expressing, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Cell Culture

    Cytokine expression in BMDC. GM-CSF-induced BMDC were separated by MACS with anti-CD11c microbeads. IFNγ, FIZZ1 and MCP-1 were detected in two distinct cell populations. *P

    Journal: PLoS ONE

    Article Title: The In Vivo Fibrotic Role of FIZZ1 in Pulmonary Fibrosis

    doi: 10.1371/journal.pone.0088362

    Figure Lengend Snippet: Cytokine expression in BMDC. GM-CSF-induced BMDC were separated by MACS with anti-CD11c microbeads. IFNγ, FIZZ1 and MCP-1 were detected in two distinct cell populations. *P

    Article Snippet: The following primary antibodies were used: anti-rat FIZZ1 (Alexis Biochemicals, San Diego, CA), anti-α-SMA (Sigma), anti-rat or mouse type I collagen (Biodesign International, Saco, ME), and HRP-conjugated anti GAPDH (Abcam, Cambridge, MA).

    Techniques: Expressing, Magnetic Cell Separation

    AdFIZZ1 effects on lung fibrosis. The lung FIZZ1 mRNA (A) at the indicated time points and protein at day 21 (B) after AdFIZZ1 endotracheal administration alone, or with BLM injection (C) were analyzed by qPCR. The results were shown as mean ± SE of triplicate samples or animals. Type I collagen production in the lungs was analyzed at day 7 after indicated treatments with 200 ng lung tissue lysates by ELISA (D). Data were shown as mean ± SE with 5 samples in each group. Lung α-SMA mRNA at day 7 (E) and protein (F) after AdFIZZ1 injection with BLM or PBS were analyzed by qPCR or Western blotting, respectively. A representative blot was shown in (F).*P

    Journal: PLoS ONE

    Article Title: The In Vivo Fibrotic Role of FIZZ1 in Pulmonary Fibrosis

    doi: 10.1371/journal.pone.0088362

    Figure Lengend Snippet: AdFIZZ1 effects on lung fibrosis. The lung FIZZ1 mRNA (A) at the indicated time points and protein at day 21 (B) after AdFIZZ1 endotracheal administration alone, or with BLM injection (C) were analyzed by qPCR. The results were shown as mean ± SE of triplicate samples or animals. Type I collagen production in the lungs was analyzed at day 7 after indicated treatments with 200 ng lung tissue lysates by ELISA (D). Data were shown as mean ± SE with 5 samples in each group. Lung α-SMA mRNA at day 7 (E) and protein (F) after AdFIZZ1 injection with BLM or PBS were analyzed by qPCR or Western blotting, respectively. A representative blot was shown in (F).*P

    Article Snippet: The following primary antibodies were used: anti-rat FIZZ1 (Alexis Biochemicals, San Diego, CA), anti-α-SMA (Sigma), anti-rat or mouse type I collagen (Biodesign International, Saco, ME), and HRP-conjugated anti GAPDH (Abcam, Cambridge, MA).

    Techniques: Injection, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot

    Generation of FIZZ1 KO mice. (A) Gene targeting strategy and restriction map of FIZZ1 gene. The diagram showed the wild type FIZZ1 gene allele and the gene targeted allele. The black boxes E1–E4 represent the four exons of FIZZ1 gene. All four exons of the FIZZ1 gene were replaced by a Neo gene cassette. (B) Southern blotting analysis of an ES cell clone with homologous recombination. A 3′ probe (shown in A) was used to detect 26 kb WT and 10 kb KO alleles. (C) PCR genotype using mouse tail DNA from wild type (“WT”), FIZZ1 heterozygote (“Het”) and homozygous knockout (“KO”) mice. The PCR fragment for WT was 525 bp, and KO was 351 bp. “Lad” referred to 100 bp ladder.

    Journal: PLoS ONE

    Article Title: The In Vivo Fibrotic Role of FIZZ1 in Pulmonary Fibrosis

    doi: 10.1371/journal.pone.0088362

    Figure Lengend Snippet: Generation of FIZZ1 KO mice. (A) Gene targeting strategy and restriction map of FIZZ1 gene. The diagram showed the wild type FIZZ1 gene allele and the gene targeted allele. The black boxes E1–E4 represent the four exons of FIZZ1 gene. All four exons of the FIZZ1 gene were replaced by a Neo gene cassette. (B) Southern blotting analysis of an ES cell clone with homologous recombination. A 3′ probe (shown in A) was used to detect 26 kb WT and 10 kb KO alleles. (C) PCR genotype using mouse tail DNA from wild type (“WT”), FIZZ1 heterozygote (“Het”) and homozygous knockout (“KO”) mice. The PCR fragment for WT was 525 bp, and KO was 351 bp. “Lad” referred to 100 bp ladder.

    Article Snippet: The following primary antibodies were used: anti-rat FIZZ1 (Alexis Biochemicals, San Diego, CA), anti-α-SMA (Sigma), anti-rat or mouse type I collagen (Biodesign International, Saco, ME), and HRP-conjugated anti GAPDH (Abcam, Cambridge, MA).

    Techniques: Mouse Assay, Southern Blot, Homologous Recombination, Polymerase Chain Reaction, Knock-Out

    Construction of AdFIZZ1. AdCMVFIZZ1.dlE3 was generated by inserting a 500-LoxP recombination between shuttle vector and the cAd5-deltaE3.LoxP cosmid. Rat FIZZ1 cDNA was under CMV promoter. “ITR” referred to the Ad5ITR and Packaging signal.

    Journal: PLoS ONE

    Article Title: The In Vivo Fibrotic Role of FIZZ1 in Pulmonary Fibrosis

    doi: 10.1371/journal.pone.0088362

    Figure Lengend Snippet: Construction of AdFIZZ1. AdCMVFIZZ1.dlE3 was generated by inserting a 500-LoxP recombination between shuttle vector and the cAd5-deltaE3.LoxP cosmid. Rat FIZZ1 cDNA was under CMV promoter. “ITR” referred to the Ad5ITR and Packaging signal.

    Article Snippet: The following primary antibodies were used: anti-rat FIZZ1 (Alexis Biochemicals, San Diego, CA), anti-α-SMA (Sigma), anti-rat or mouse type I collagen (Biodesign International, Saco, ME), and HRP-conjugated anti GAPDH (Abcam, Cambridge, MA).

    Techniques: Generated, Plasmid Preparation

    Effects of FIZZ1 deficiency on BLM-induced pulmonary fibrosis. WT control or FIZZ1 KO mice were treated with PBS (CON) or BLM as indicated. The lungs were harvested at day 21, and analyzed for lung HYP content (A). The values were expressed as percentages of their respective PBS control. Data were shown as mean ± SE with 5 mice in each group. *P

    Journal: PLoS ONE

    Article Title: The In Vivo Fibrotic Role of FIZZ1 in Pulmonary Fibrosis

    doi: 10.1371/journal.pone.0088362

    Figure Lengend Snippet: Effects of FIZZ1 deficiency on BLM-induced pulmonary fibrosis. WT control or FIZZ1 KO mice were treated with PBS (CON) or BLM as indicated. The lungs were harvested at day 21, and analyzed for lung HYP content (A). The values were expressed as percentages of their respective PBS control. Data were shown as mean ± SE with 5 mice in each group. *P

    Article Snippet: The following primary antibodies were used: anti-rat FIZZ1 (Alexis Biochemicals, San Diego, CA), anti-α-SMA (Sigma), anti-rat or mouse type I collagen (Biodesign International, Saco, ME), and HRP-conjugated anti GAPDH (Abcam, Cambridge, MA).

    Techniques: Mouse Assay

    Effect of FIZZ1 treatment on lung fibroblast-expressed Notch1 signaling components. Normal murine lung fibroblasts were treated with the indicated doses of rmFIZZ1 for 1 hour, or as indicated. A : Cell lysates (20 μg protein each) were analyzed

    Journal: The American Journal of Pathology

    Article Title: Notch1 Signaling in FIZZ1 Induction of Myofibroblast Differentiation

    doi: 10.2353/ajpath.2009.080618

    Figure Lengend Snippet: Effect of FIZZ1 treatment on lung fibroblast-expressed Notch1 signaling components. Normal murine lung fibroblasts were treated with the indicated doses of rmFIZZ1 for 1 hour, or as indicated. A : Cell lysates (20 μg protein each) were analyzed

    Article Snippet: Where indicated, recombinant mouse FIZZ1 (rmFIZZ1; Leinco Technologies, Inc., St. Louis, MO) were added at the indicated doses.

    Techniques:

    Effects of Notch deficiency on lung cytokine and FIZZ1 gene expression in BLM-induced pulmonary fibrosis. FX KO mice were maintained with (Notch-sufficient) or without (Notch-deficient) dietary fucose supplementation, and then treated with saline (SAL)

    Journal: The American Journal of Pathology

    Article Title: Notch1 Signaling in FIZZ1 Induction of Myofibroblast Differentiation

    doi: 10.2353/ajpath.2009.080618

    Figure Lengend Snippet: Effects of Notch deficiency on lung cytokine and FIZZ1 gene expression in BLM-induced pulmonary fibrosis. FX KO mice were maintained with (Notch-sufficient) or without (Notch-deficient) dietary fucose supplementation, and then treated with saline (SAL)

    Article Snippet: Where indicated, recombinant mouse FIZZ1 (rmFIZZ1; Leinco Technologies, Inc., St. Louis, MO) were added at the indicated doses.

    Techniques: Expressing, Mouse Assay

    Schematic illustration of the proposed mechanism by which Notch1/Jagged1 mediates FIZZ1 induction of α-SMA gene expression. FIZZ1 is primarily secreted by alveolar epithelial cells in the lung, and activates Jagged1 expression through unknown

    Journal: The American Journal of Pathology

    Article Title: Notch1 Signaling in FIZZ1 Induction of Myofibroblast Differentiation

    doi: 10.2353/ajpath.2009.080618

    Figure Lengend Snippet: Schematic illustration of the proposed mechanism by which Notch1/Jagged1 mediates FIZZ1 induction of α-SMA gene expression. FIZZ1 is primarily secreted by alveolar epithelial cells in the lung, and activates Jagged1 expression through unknown

    Article Snippet: Where indicated, recombinant mouse FIZZ1 (rmFIZZ1; Leinco Technologies, Inc., St. Louis, MO) were added at the indicated doses.

    Techniques: Expressing

    Effect of Notch deficiency on FIZZ1 induction of α-SMA in isolated lung fibroblasts. Murine lung fibroblasts were isolated from FX KO mice and were cultured in the presence (Notch-sufficient, indicated with +) or absence (Notch-deficient,

    Journal: The American Journal of Pathology

    Article Title: Notch1 Signaling in FIZZ1 Induction of Myofibroblast Differentiation

    doi: 10.2353/ajpath.2009.080618

    Figure Lengend Snippet: Effect of Notch deficiency on FIZZ1 induction of α-SMA in isolated lung fibroblasts. Murine lung fibroblasts were isolated from FX KO mice and were cultured in the presence (Notch-sufficient, indicated with +) or absence (Notch-deficient,

    Article Snippet: Where indicated, recombinant mouse FIZZ1 (rmFIZZ1; Leinco Technologies, Inc., St. Louis, MO) were added at the indicated doses.

    Techniques: Isolation, Mouse Assay, Cell Culture

    Role of Jagged1 in FIZZ1 induction of α-SMA expression. A : Fibroblasts were transfected as indicated with either Jagged1 expression plasmid or its empty vector control, pcDNA3.1, and then treated with buffer only or FIZZ1. The cells were then

    Journal: The American Journal of Pathology

    Article Title: Notch1 Signaling in FIZZ1 Induction of Myofibroblast Differentiation

    doi: 10.2353/ajpath.2009.080618

    Figure Lengend Snippet: Role of Jagged1 in FIZZ1 induction of α-SMA expression. A : Fibroblasts were transfected as indicated with either Jagged1 expression plasmid or its empty vector control, pcDNA3.1, and then treated with buffer only or FIZZ1. The cells were then

    Article Snippet: Where indicated, recombinant mouse FIZZ1 (rmFIZZ1; Leinco Technologies, Inc., St. Louis, MO) were added at the indicated doses.

    Techniques: Expressing, Transfection, Plasmid Preparation

    Photomicrographs of M2 microglia following GCM stimulation of glioma cells in vitro and in vivo. (a) The human microglia HMC3 cells were treated with GCM from LN229, LN229R, HG7 and HG7R cells. (b) HMC3 were treated with GCM from LN229-Scr, LN229-OE, HG7-Scr and HG7-OE cells. (c) BV-2 cells were treated with GCM from GL261, GL261R, GL261-Scr and GL261-OE cells. CD68, CD163, CD204 and CD206 were stained in red, while Arg1, Mrc1, Fizz1 and Cd163 were stained in green. Nuclei were stained with DAPI (blue). (d) IHC assays were performed to detect Arg1, Mrc1, Fizz1 and Cd163 in xenograft gliomas formed by GL261-Scr and GL261-OE cells. All images were taken microscopically (20× or 40×). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Journal: EBioMedicine

    Article Title: DNA damage repair alterations modulate M2 polarization of microglia to remodel the tumor microenvironment via the p53-mediated MDK expression in glioma

    doi: 10.1016/j.ebiom.2019.01.067

    Figure Lengend Snippet: Photomicrographs of M2 microglia following GCM stimulation of glioma cells in vitro and in vivo. (a) The human microglia HMC3 cells were treated with GCM from LN229, LN229R, HG7 and HG7R cells. (b) HMC3 were treated with GCM from LN229-Scr, LN229-OE, HG7-Scr and HG7-OE cells. (c) BV-2 cells were treated with GCM from GL261, GL261R, GL261-Scr and GL261-OE cells. CD68, CD163, CD204 and CD206 were stained in red, while Arg1, Mrc1, Fizz1 and Cd163 were stained in green. Nuclei were stained with DAPI (blue). (d) IHC assays were performed to detect Arg1, Mrc1, Fizz1 and Cd163 in xenograft gliomas formed by GL261-Scr and GL261-OE cells. All images were taken microscopically (20× or 40×). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Briefly, the slices were incubated with primary antibody rabbit anti-C5 (1:100, ABclonal, A8104), rabbit anti-IL6 (1:200, ABclonal, A0286), mouse anti-TNFSF4 (1:200, Abcam, ab89896), rabbit anti-TNFSF4 (1:200, ThermoFisher, #MA5–17910), rabbit anti-SAA1 (1:200, Abcam, ab207445), rabbit anti-SAA1 (1:200, Abcam, ab199030), rabbit anti-MDK (1:200, Abcam, ab170820), mouse anti-VEGFA (1:200, Abcam, ab1316), rabbit anti-CD68 (1:200, Abcam, ab125212), rabbit anti-CD163 (1:200, Proteintech, 16,646–1-AP), rabbit anti-CD204 (1:200, Abcam, ab123946), rabbit anti-CD206 (1:200, Abcam, ab64693), mouse anti-Arg1 (1:200, Abcam, ab239731), rabbit anti-Fizz1 (1:200, Abcam, ab39626) and rabbit anti-Mrc1 (1:200, Abcam, ab64693) for 12 h at 4 °C.

    Techniques: In Vitro, In Vivo, Staining, Immunohistochemistry

    FIZZ1 and YM1 expression in lung epithelial cells. Allergic lung inflammation was induced in RAG2 −/− and γ c −/− mice as mentioned in Fig. 1. FIZZ1 and YM1 expression was analyzed in serial sections of mouse lungs by immunohistochemistry. Photomicrographs (40X magnification) of YM1 (panels a-d) and FIZZ1 (panels e-h) expression in epithelial cells in representative lung sections are shown. (B) The number of YM1+ or FIZZ1+ airways in each group of mice was counted. Data represented as number of airways ± SEM. *p

    Journal: PLoS ONE

    Article Title: Absence of the Common Gamma Chain (?c), a Critical Component of the Type I IL-4 Receptor, Increases the Severity of Allergic Lung Inflammation

    doi: 10.1371/journal.pone.0071344

    Figure Lengend Snippet: FIZZ1 and YM1 expression in lung epithelial cells. Allergic lung inflammation was induced in RAG2 −/− and γ c −/− mice as mentioned in Fig. 1. FIZZ1 and YM1 expression was analyzed in serial sections of mouse lungs by immunohistochemistry. Photomicrographs (40X magnification) of YM1 (panels a-d) and FIZZ1 (panels e-h) expression in epithelial cells in representative lung sections are shown. (B) The number of YM1+ or FIZZ1+ airways in each group of mice was counted. Data represented as number of airways ± SEM. *p

    Article Snippet: For immunohistochemistry, deparaffinized sections were incubated with 10% goat serum and stained with a 1∶100 dilution of rabbit anti-mouse FIZZ1 (Abcam, Cambridge, MA) or 1∶100 dilution of rabbit anti-mouse YM1 (Stem Cell Technologies, Vancouver, Canada).

    Techniques: Expressing, Mouse Assay, Immunohistochemistry

    FIZZ1 and YM1 expression in macrophages. Allergic lung disease was induced in RAG2 −/− and γ c −/− mice as shown in Figure 1 . Serial sections of mouse lungs were stained for FIZZ1 and YM1 by immunohistochemistry. Photomicrographs (100X magnification) of YM1 (panels a-d) and FIZZ1 (panels e-h) expression in macrophages in representative lung sections are shown. YM1 + macrophages are indicated by arrows. (B) BAL cells from RAG2 −/− and γcxRAG2 −/− mice immunized and challenged with OVA were collected and analyzed by FACS. Cells were labeled with a fluorochrome-conjugated antibody to CD11b, stained with an antibody to YM1, followed by a secondary antibody conjugated to Alexa Fluor 647 (solid histogram). Secondary antibody staining alone was used as control (dashed histogram). Macrophages were gated based on forward by side scatter and then on CD11b expression. MFI = Mean Fluorescence Intensity of YM1 expression. (C) The average MFI of YM1 staining in macrophages from RAG2 −/− and γcxRAG2 −/− mice from different experiments is shown. *p

    Journal: PLoS ONE

    Article Title: Absence of the Common Gamma Chain (?c), a Critical Component of the Type I IL-4 Receptor, Increases the Severity of Allergic Lung Inflammation

    doi: 10.1371/journal.pone.0071344

    Figure Lengend Snippet: FIZZ1 and YM1 expression in macrophages. Allergic lung disease was induced in RAG2 −/− and γ c −/− mice as shown in Figure 1 . Serial sections of mouse lungs were stained for FIZZ1 and YM1 by immunohistochemistry. Photomicrographs (100X magnification) of YM1 (panels a-d) and FIZZ1 (panels e-h) expression in macrophages in representative lung sections are shown. YM1 + macrophages are indicated by arrows. (B) BAL cells from RAG2 −/− and γcxRAG2 −/− mice immunized and challenged with OVA were collected and analyzed by FACS. Cells were labeled with a fluorochrome-conjugated antibody to CD11b, stained with an antibody to YM1, followed by a secondary antibody conjugated to Alexa Fluor 647 (solid histogram). Secondary antibody staining alone was used as control (dashed histogram). Macrophages were gated based on forward by side scatter and then on CD11b expression. MFI = Mean Fluorescence Intensity of YM1 expression. (C) The average MFI of YM1 staining in macrophages from RAG2 −/− and γcxRAG2 −/− mice from different experiments is shown. *p

    Article Snippet: For immunohistochemistry, deparaffinized sections were incubated with 10% goat serum and stained with a 1∶100 dilution of rabbit anti-mouse FIZZ1 (Abcam, Cambridge, MA) or 1∶100 dilution of rabbit anti-mouse YM1 (Stem Cell Technologies, Vancouver, Canada).

    Techniques: Expressing, Mouse Assay, Staining, Immunohistochemistry, FACS, Labeling, Fluorescence

    Fizz1- and Ym1-expressing macrophages in wound tissue of IL-10 −/− and control mice. Day 5 wounds of control ( A , B ) and IL-10 −/− mice ( C , D ) stained for Fizz1 ( A , C ) or Ym1 ( B , D ); numerous mononuclear cells within the granulation tissue of control and mutant mice stain positive for Fizz1 or Ym1. E: Double labeling for F4/80 (green) and Fizz1 or Ym1 (red) in granulation tissue of IL-10 −/− mice demonstrates Fizz1 and Ym1 expression by macrophages. Scale bars = 100 μm. Original magnifications, ×400 ( E ).

    Journal: The American Journal of Pathology

    Article Title: Accelerated Wound Closure in Mice Deficient for Interleukin-10

    doi: 10.2353/ajpath.2007.060370

    Figure Lengend Snippet: Fizz1- and Ym1-expressing macrophages in wound tissue of IL-10 −/− and control mice. Day 5 wounds of control ( A , B ) and IL-10 −/− mice ( C , D ) stained for Fizz1 ( A , C ) or Ym1 ( B , D ); numerous mononuclear cells within the granulation tissue of control and mutant mice stain positive for Fizz1 or Ym1. E: Double labeling for F4/80 (green) and Fizz1 or Ym1 (red) in granulation tissue of IL-10 −/− mice demonstrates Fizz1 and Ym1 expression by macrophages. Scale bars = 100 μm. Original magnifications, ×400 ( E ).

    Article Snippet: For Fizz1 and Ym1 detection, cryosections were incubated for 1 hour at room temperature with rabbit anti-Fizz1 (Peprotech, London, UK) or goat anti-Ym1 (R & R Systems, Wiesbaden, Germany) antibody (1:50); bound primary antibodies were detected using a peroxidase-conjugated anti-rabbit or anti-goat antibody (DakoCytomation), respectively.

    Techniques: Expressing, Mouse Assay, Staining, Mutagenesis, Labeling

    Lung macrophages prominently express M2 proteins following IL-4-treatment in intratracheal LPS-exposed WT mice. A : mRNA expression for the M2 markers Arg1 , Fizz1 , and Ym1 quantified in the whole lung at days 4 and 6 among intratracheal H 2 O- or LPS-exposed

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Enhanced resolution of experimental ARDS through IL-4-mediated lung macrophage reprogramming

    doi: 10.1152/ajplung.00419.2015

    Figure Lengend Snippet: Lung macrophages prominently express M2 proteins following IL-4-treatment in intratracheal LPS-exposed WT mice. A : mRNA expression for the M2 markers Arg1 , Fizz1 , and Ym1 quantified in the whole lung at days 4 and 6 among intratracheal H 2 O- or LPS-exposed

    Article Snippet: Membranes were blocked in SuperBlock T20 blocking buffer (Pierce) for 30 min at room temperature and then incubated overnight at 4°C with polyclonal rabbit anti-mouse Ym1 (STEMCELL Technologies), rabbit anti-mouse FIZZ1 (Peprotech), mouse anti-mouse Arg1 (BD Transduction Laboratories), or rabbit anti-mouse β-actin (Abcam) antibodies diluted 1:1,500 in blocking solution.

    Techniques: Mouse Assay, Expressing