fluorochrome labeled anti ifn γ  (Thermo Fisher)


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    Name:
    IFN gamma Antibody 22H17L20
    Description:
    IFN gamma Monoclonal Antibody for Western Blot
    Catalog Number:
    701121
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
    Applications:
    Build Your Own Immunoassay|Cell Analysis|ELISA|ELISAs for Cell Biology & Signal Transduction|Protein Assays and Analysis|Protein Biology|Ready-To-Use Immunoassay|Western Blot Detection|Western Blotting
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    Structured Review

    Thermo Fisher fluorochrome labeled anti ifn γ
    <t>IFN-γ</t> and IL-4 production of CD3/CD28 costimulated naive CD4 + T cells from rheumatoid arthritis patients ( n = 9) differentiated during (a) 5-day cultures or (b) 10-day cultures in the presence of IL-7, of IL-4, or of IL-7 and IL-4. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate restimulation for 24 hours. * P
    IFN gamma Monoclonal Antibody for Western Blot
    https://www.bioz.com/result/fluorochrome labeled anti ifn γ/product/Thermo Fisher
    Average 96 stars, based on 1 article reviews
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    1) Product Images from "Differentiation of naive CD4+ T cells towards T helper 2 cells is not impaired in rheumatoid arthritis patients"

    Article Title: Differentiation of naive CD4+ T cells towards T helper 2 cells is not impaired in rheumatoid arthritis patients

    Journal: Arthritis Research & Therapy

    doi:

    IFN-γ and IL-4 production of CD3/CD28 costimulated naive CD4 + T cells from rheumatoid arthritis patients ( n = 9) differentiated during (a) 5-day cultures or (b) 10-day cultures in the presence of IL-7, of IL-4, or of IL-7 and IL-4. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate restimulation for 24 hours. * P
    Figure Legend Snippet: IFN-γ and IL-4 production of CD3/CD28 costimulated naive CD4 + T cells from rheumatoid arthritis patients ( n = 9) differentiated during (a) 5-day cultures or (b) 10-day cultures in the presence of IL-7, of IL-4, or of IL-7 and IL-4. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate restimulation for 24 hours. * P

    Techniques Used:

    Intracellular detection of IFN-γ and IL-4 by FACS analysis. (a) A representative of seven rheumatoid arthritis (RA) patients of which mean values are shown in (b) . Cytokine production of the CD3/CD28 costimulated CD4 + T cells was assessed after 10 days of culture in the absence or presence of IL-7, of IL-4 or of their combination. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate stimulation for 6 hours in the presence of Brefeldin A. Means ± standard errors of the mean of percentages from cells of RA patients that produce IFN-γ but no IL-4 (Th1) and that produce IL-4 but no IFN-γ (Th2) upon differentiation are shown. * P
    Figure Legend Snippet: Intracellular detection of IFN-γ and IL-4 by FACS analysis. (a) A representative of seven rheumatoid arthritis (RA) patients of which mean values are shown in (b) . Cytokine production of the CD3/CD28 costimulated CD4 + T cells was assessed after 10 days of culture in the absence or presence of IL-7, of IL-4 or of their combination. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate stimulation for 6 hours in the presence of Brefeldin A. Means ± standard errors of the mean of percentages from cells of RA patients that produce IFN-γ but no IL-4 (Th1) and that produce IL-4 but no IFN-γ (Th2) upon differentiation are shown. * P

    Techniques Used: FACS

    IFN-γ and IL-4 production of CD3/CD28 costimulated naive CD4 + T cells from rheumatoid arthritis patients ( n = 9) stimulated for 5 days in the presence of IL-7. Because of exogenously added IL-4, only the production of IFN-γ is shown for cells cultured in the presence of IL-4 or of IL-7 and IL-4. * P
    Figure Legend Snippet: IFN-γ and IL-4 production of CD3/CD28 costimulated naive CD4 + T cells from rheumatoid arthritis patients ( n = 9) stimulated for 5 days in the presence of IL-7. Because of exogenously added IL-4, only the production of IFN-γ is shown for cells cultured in the presence of IL-4 or of IL-7 and IL-4. * P

    Techniques Used: Cell Culture

    The production of IFN-γ and IL-4 by CD3/CD28 costimulated naive CD4 + T cells from healthy controls ( n = 9, white bars) and rheumatoid arthritis (RA) patients ( n = 9, black bars) differentiated during 10 days of culture in the presence of IL-7, of IL-4 or of IL-7 and IL-4. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate restimulation for 24 hours. Values are expressed as percentages of cytokine levels produced in the absence of exogenously added cytokines (control values set at 100% for RA patients and controls, respectively, were 47.1 ± 12 ng/ml versus 37.1 ± 5.7 ng/ml for IFN-γ, and 0.89 ± 0.21 ng/ml versus 0.95 ± 0.15 ng/ml for IL-4; not significantly different). # P
    Figure Legend Snippet: The production of IFN-γ and IL-4 by CD3/CD28 costimulated naive CD4 + T cells from healthy controls ( n = 9, white bars) and rheumatoid arthritis (RA) patients ( n = 9, black bars) differentiated during 10 days of culture in the presence of IL-7, of IL-4 or of IL-7 and IL-4. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate restimulation for 24 hours. Values are expressed as percentages of cytokine levels produced in the absence of exogenously added cytokines (control values set at 100% for RA patients and controls, respectively, were 47.1 ± 12 ng/ml versus 37.1 ± 5.7 ng/ml for IFN-γ, and 0.89 ± 0.21 ng/ml versus 0.95 ± 0.15 ng/ml for IL-4; not significantly different). # P

    Techniques Used: Produced

    2) Product Images from "DGAT1 inhibits retinol-dependent regulatory T cell formation and mediates autoimmune encephalomyelitis"

    Article Title: DGAT1 inhibits retinol-dependent regulatory T cell formation and mediates autoimmune encephalomyelitis

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

    doi: 10.1073/pnas.1817669116

    Effect of DGAT1 deficiency on numbers of IFN-γ– or IL-17–producing CD4 + T cells in dLN and CNS. Male WT and DGAT1 KO mice ( n = 5 per group) were induced to develop EAE. At day 17 postimmunization, mice were killed. dLN cells ( Upper ) or CNS mononuclear cells ( Lower ) were restimulated for 3 d with MOG 35–55 , and PMA/ionomycin and protein transport inhibitor were added for the last 5 h of the culture period. Cells were then stained with mAbs to surface markers and intracellular cytokines and analyzed by flow cytometry. Viable lymphocytes were gated as NK1.1 − CD4 + and evaluated for IFN-γ and IL-17 expression. ( A ) Representative FACS plots indicate the percentage of CD4 + T cells that stained positive for IFN-γ or IL-17 in WT and DGAT1 KO dLNs and CNS. ( B ) Scatter plots indicate the percentage of IFN-γ + ( Left ) and IL-17 + ( Right ) CD4 + T cells in WT and DGAT1 KO tissues. Each symbol represents an individual mouse; the bar depicts the mean ± SEM. None of the differences achieved statistical significance.
    Figure Legend Snippet: Effect of DGAT1 deficiency on numbers of IFN-γ– or IL-17–producing CD4 + T cells in dLN and CNS. Male WT and DGAT1 KO mice ( n = 5 per group) were induced to develop EAE. At day 17 postimmunization, mice were killed. dLN cells ( Upper ) or CNS mononuclear cells ( Lower ) were restimulated for 3 d with MOG 35–55 , and PMA/ionomycin and protein transport inhibitor were added for the last 5 h of the culture period. Cells were then stained with mAbs to surface markers and intracellular cytokines and analyzed by flow cytometry. Viable lymphocytes were gated as NK1.1 − CD4 + and evaluated for IFN-γ and IL-17 expression. ( A ) Representative FACS plots indicate the percentage of CD4 + T cells that stained positive for IFN-γ or IL-17 in WT and DGAT1 KO dLNs and CNS. ( B ) Scatter plots indicate the percentage of IFN-γ + ( Left ) and IL-17 + ( Right ) CD4 + T cells in WT and DGAT1 KO tissues. Each symbol represents an individual mouse; the bar depicts the mean ± SEM. None of the differences achieved statistical significance.

    Techniques Used: Mouse Assay, Staining, Flow Cytometry, Cytometry, Expressing, FACS

    3) Product Images from "Mycophenolic acid derivative 118 improves outcome of skin grafts by suppressing IL-17 production"

    Article Title: Mycophenolic acid derivative 118 improves outcome of skin grafts by suppressing IL-17 production

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2013.14

    118 did not strongly affect Th17 cell differentiation in vitro . Naïve CD4 + T cells were cultured under Th17 polarizing conditions and analyzed for IFN-γ and IL-17 expression by intracellular staining. The numbers indicate the percentages
    Figure Legend Snippet: 118 did not strongly affect Th17 cell differentiation in vitro . Naïve CD4 + T cells were cultured under Th17 polarizing conditions and analyzed for IFN-γ and IL-17 expression by intracellular staining. The numbers indicate the percentages

    Techniques Used: Cell Differentiation, In Vitro, Cell Culture, Expressing, Staining

    4) Product Images from "Identification and Evaluation of Novel Protective Antigens for the Development of a Candidate Tuberculosis Subunit Vaccine"

    Article Title: Identification and Evaluation of Novel Protective Antigens for the Development of a Candidate Tuberculosis Subunit Vaccine

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00014-18

    Measurement of the breadth and strength of the immune response induced after administration of the different ChAdOx1 vaccines. The antigen-specific IFN-γ immune responses in BALB/c and C57BL/6 mice were measured at 2 weeks postimmunization. (a and b) ChAdOx1.PPE15 vaccination; (c and d) ChAdOx1.PPE51 vaccination; (e and f) ChAdOx1.PE3 vaccination; (g and h) ChAdOx1.PE12 vaccination. Splenocytes from three vaccinated animals were stimulated with pools of 10 15-mer peptides overlapping by 11 amino acids. Each dot represents one animal, and the lines represent the median response. Data are representative of those from two independent experiments.
    Figure Legend Snippet: Measurement of the breadth and strength of the immune response induced after administration of the different ChAdOx1 vaccines. The antigen-specific IFN-γ immune responses in BALB/c and C57BL/6 mice were measured at 2 weeks postimmunization. (a and b) ChAdOx1.PPE15 vaccination; (c and d) ChAdOx1.PPE51 vaccination; (e and f) ChAdOx1.PE3 vaccination; (g and h) ChAdOx1.PE12 vaccination. Splenocytes from three vaccinated animals were stimulated with pools of 10 15-mer peptides overlapping by 11 amino acids. Each dot represents one animal, and the lines represent the median response. Data are representative of those from two independent experiments.

    Techniques Used: Mouse Assay

    Antigen-specific responses after BCG and ChAdOx1 immunizations. (a and b) Splenocytes from BCG-vaccinated BALB/c (a) and C57BL/6 (b) mice were stimulated with PPE51, PPE15, PE3, or PE12, and IFN-γ production was measured at different time points postvaccination using an ELISpot assay. Stimulations with Ag85A and PPD-T were used as positive controls. Each symbol represents the median response (in number of spot-forming units [SFU]/10 6 cells) for three animals, and the error bars represent the range of values. (c to f) CB6F1 mice were vaccinated with a single intranasal administration of ChAdOx1 expressing the four selected antigens; the percentage of antigen-specific CD4 + or CD8 + T cells expressing IFN-γ, TNF-α, and IL-2 was measured in the lungs (c and d) and spleen (e and f). The bars represent the median response of five animals, and error bars indicate the interquartile range. Data are representative of those from two independent experiments.
    Figure Legend Snippet: Antigen-specific responses after BCG and ChAdOx1 immunizations. (a and b) Splenocytes from BCG-vaccinated BALB/c (a) and C57BL/6 (b) mice were stimulated with PPE51, PPE15, PE3, or PE12, and IFN-γ production was measured at different time points postvaccination using an ELISpot assay. Stimulations with Ag85A and PPD-T were used as positive controls. Each symbol represents the median response (in number of spot-forming units [SFU]/10 6 cells) for three animals, and the error bars represent the range of values. (c to f) CB6F1 mice were vaccinated with a single intranasal administration of ChAdOx1 expressing the four selected antigens; the percentage of antigen-specific CD4 + or CD8 + T cells expressing IFN-γ, TNF-α, and IL-2 was measured in the lungs (c and d) and spleen (e and f). The bars represent the median response of five animals, and error bars indicate the interquartile range. Data are representative of those from two independent experiments.

    Techniques Used: Mouse Assay, Enzyme-linked Immunospot, Expressing

    5) Product Images from "Host innate recognition of an intestinal bacterial pathogen induces TRIF-dependent protective immunity"

    Article Title: Host innate recognition of an intestinal bacterial pathogen induces TRIF-dependent protective immunity

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20110547

    TRIF-mediated IFN-β induction by macrophages elicits IFN-γ production from NK cells which further enhances the bactericidal function of the macrophages. (A) Peritoneal macrophages isolated from WT and Trif LPS2 mice were stimulated with poly I:C for 6 h in the presence or absence of anti-IFNAR1 antibody. After washing out poly I:C, macrophages were co-cultured with WT splenic NK cells for 24 h in the presence or absence of neutralizing anti–IFN-γ antibody. After excluding NK cells, the remaining macrophages were subjected to bactericidal assay. (B) ELISA measurement of IFN-γ production from WT NK cells after co-culturing with poly I:C–stimulated macrophages in the presence or absence of anti-IFNAR1 antibody. Combined data from three independent experiments ( n = 6 each; *, P
    Figure Legend Snippet: TRIF-mediated IFN-β induction by macrophages elicits IFN-γ production from NK cells which further enhances the bactericidal function of the macrophages. (A) Peritoneal macrophages isolated from WT and Trif LPS2 mice were stimulated with poly I:C for 6 h in the presence or absence of anti-IFNAR1 antibody. After washing out poly I:C, macrophages were co-cultured with WT splenic NK cells for 24 h in the presence or absence of neutralizing anti–IFN-γ antibody. After excluding NK cells, the remaining macrophages were subjected to bactericidal assay. (B) ELISA measurement of IFN-γ production from WT NK cells after co-culturing with poly I:C–stimulated macrophages in the presence or absence of anti-IFNAR1 antibody. Combined data from three independent experiments ( n = 6 each; *, P

    Techniques Used: Isolation, Mouse Assay, Cell Culture, Serum Bactericidal Assay, Enzyme-linked Immunosorbent Assay

    6) Product Images from "HMPL-004 (Andrographis paniculata extract) prevents development of murine colitis by inhibiting T cell proliferation and TH1/TH17 responses"

    Article Title: HMPL-004 (Andrographis paniculata extract) prevents development of murine colitis by inhibiting T cell proliferation and TH1/TH17 responses

    Journal: Inflammatory bowel diseases

    doi: 10.1002/ibd.22983

    HMPL-004 inhibits the early T H 17 polarization in vivo ( A ) Total numbers of mononuclear cells in spleen, and LP of HMPL-004 and MC treated mice one week after T cell transfer (n=11/group for spleens, n=3/group for LPMC). ( B ) Percentage of CD4 + splenocytes of HMPL-004 and MC treated mice. (n=11/group). ( C ) Representative dot blots of intracellular IFN-γ and IL-17 staining of PMA + Ionomycin stimulated splenocytes isolated from HMPL-004 and MC treated mice one week after T cell transfer. Gated on CD4 + T cells. ( D ) Percentages of CD4 + IL-17 + or CD4 + IFN-γ + splenocytes of HMPL-004 and MC treated mice. (n=7–8/group). All data represent the mean ± SD. Statistical significance was determined by Student’s t-test (A, B) or Wilcoxon rank sum test (D). *, p
    Figure Legend Snippet: HMPL-004 inhibits the early T H 17 polarization in vivo ( A ) Total numbers of mononuclear cells in spleen, and LP of HMPL-004 and MC treated mice one week after T cell transfer (n=11/group for spleens, n=3/group for LPMC). ( B ) Percentage of CD4 + splenocytes of HMPL-004 and MC treated mice. (n=11/group). ( C ) Representative dot blots of intracellular IFN-γ and IL-17 staining of PMA + Ionomycin stimulated splenocytes isolated from HMPL-004 and MC treated mice one week after T cell transfer. Gated on CD4 + T cells. ( D ) Percentages of CD4 + IL-17 + or CD4 + IFN-γ + splenocytes of HMPL-004 and MC treated mice. (n=7–8/group). All data represent the mean ± SD. Statistical significance was determined by Student’s t-test (A, B) or Wilcoxon rank sum test (D). *, p

    Techniques Used: In Vivo, Mouse Assay, Staining, Isolation

    HMPL-004 inhibits the proliferation of CD4 + T cells in vitro CD4 + CD25 − T cells were isolated from the spleen of mice and labeled with CFSE. T cells were stimulated with anti-CD3 and anti-CD28 in the presence of HMPL-004 (1–10 µg/ml) or MC for 4 days. ( A ) The proliferation of CD4 + T cells in the presence of HMPL-004 was analyzed using flow cytometry and was compared to MC treated cells. ( B ) Secretion of IFN-γ was analyzed by ELISA. One representative experiment out of three independent experiments with similar results is shown.
    Figure Legend Snippet: HMPL-004 inhibits the proliferation of CD4 + T cells in vitro CD4 + CD25 − T cells were isolated from the spleen of mice and labeled with CFSE. T cells were stimulated with anti-CD3 and anti-CD28 in the presence of HMPL-004 (1–10 µg/ml) or MC for 4 days. ( A ) The proliferation of CD4 + T cells in the presence of HMPL-004 was analyzed using flow cytometry and was compared to MC treated cells. ( B ) Secretion of IFN-γ was analyzed by ELISA. One representative experiment out of three independent experiments with similar results is shown.

    Techniques Used: In Vitro, Isolation, Mouse Assay, Labeling, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    HMPL-004 inhibits the differentiation and/or expansion of naïve CD4 + T cells in vitro Naïve CD4 + T cells were isolated, CFSE-labeled, and stimulated under T H 0, T H 1, and T H 17 conditions in the presence of HMPL-004 (10–20 µg/ml) or MC for 5 days. ( A ) The proliferation of CD4 + T cells treated with HMPL-004 was analyzed using flow cytometry and was compared to MC treated cells. ( B ) Secretion of IFN-γ and IL-17 for T H 1 and T H 17 cells, respectively, was analyzed by ELISA. ( C ) T H 1 cells were restimulated with PMA + Ionomycin and stained for CD4 and intracellular expression of IFN-γ and IL-17. Flow cytometry plots are gated on CD4 + T cells. Dot plots show the expression of IFN-γ and IL-17 (left panels). Histograms are gated on either the IFN-γ + (middle panels) or IFN-γ − (right panels) population and show CFSE dilution. ( D ) T H 17 cells were restimulated with PMA + Ionomycin and stained for CD4 and intracellular expression of IFN-γ and IL-17. Flow cytometry plots are gated on CD4 + T cells. Dot plots show the expression of IFN-γ and IL-17 (left panels). Histograms are gated on either the IL-17 + , IL-17 − /IFN-γ − , or IFN-γ + population and show CFSE dilution. One representative experiment out of three independent experiments with similar results is shown.
    Figure Legend Snippet: HMPL-004 inhibits the differentiation and/or expansion of naïve CD4 + T cells in vitro Naïve CD4 + T cells were isolated, CFSE-labeled, and stimulated under T H 0, T H 1, and T H 17 conditions in the presence of HMPL-004 (10–20 µg/ml) or MC for 5 days. ( A ) The proliferation of CD4 + T cells treated with HMPL-004 was analyzed using flow cytometry and was compared to MC treated cells. ( B ) Secretion of IFN-γ and IL-17 for T H 1 and T H 17 cells, respectively, was analyzed by ELISA. ( C ) T H 1 cells were restimulated with PMA + Ionomycin and stained for CD4 and intracellular expression of IFN-γ and IL-17. Flow cytometry plots are gated on CD4 + T cells. Dot plots show the expression of IFN-γ and IL-17 (left panels). Histograms are gated on either the IFN-γ + (middle panels) or IFN-γ − (right panels) population and show CFSE dilution. ( D ) T H 17 cells were restimulated with PMA + Ionomycin and stained for CD4 and intracellular expression of IFN-γ and IL-17. Flow cytometry plots are gated on CD4 + T cells. Dot plots show the expression of IFN-γ and IL-17 (left panels). Histograms are gated on either the IL-17 + , IL-17 − /IFN-γ − , or IFN-γ + population and show CFSE dilution. One representative experiment out of three independent experiments with similar results is shown.

    Techniques Used: In Vitro, Isolation, Labeling, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Expressing

    HMPL-004 treatment inhibits the expression of pro-inflammatory mediators but has no effect on anti-inflammatory cytokines ( A ) TNF-α, IFN-γ, IL-22, IL-1β, and IL-10 mRNA expression was determined in cecum and colon of HMPL-004 and MC treated mice by real-time PCR. Data were normalized to the expression of β-actin mRNA. (n=5/group) ( B ) IL-6 serum concentration of HMPL-004 and MC treated mice at day of sacrifice (n=15–16/group). ( C ) TNF-α serum concentration of HMPL-004 and MC treated mice at day of sacrifice (n=13–15/group). All data represent the mean ± SD. Statistical significance was determined by Student’s t-test . *, p
    Figure Legend Snippet: HMPL-004 treatment inhibits the expression of pro-inflammatory mediators but has no effect on anti-inflammatory cytokines ( A ) TNF-α, IFN-γ, IL-22, IL-1β, and IL-10 mRNA expression was determined in cecum and colon of HMPL-004 and MC treated mice by real-time PCR. Data were normalized to the expression of β-actin mRNA. (n=5/group) ( B ) IL-6 serum concentration of HMPL-004 and MC treated mice at day of sacrifice (n=15–16/group). ( C ) TNF-α serum concentration of HMPL-004 and MC treated mice at day of sacrifice (n=13–15/group). All data represent the mean ± SD. Statistical significance was determined by Student’s t-test . *, p

    Techniques Used: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction, Concentration Assay

    HMPL-004 inhibits the early proliferation of CD4 + T cells, and T H 1 and T H 17 polarization in vivo ( A ) Total numbers of mononuclear cells in LP, MLN, and spleen of HMPL-004 and MC treated mice two weeks after T cell transfer (n=11/group for spleens, n=6/group for LPMC, n=8/group for MLN). ( B ) Percentage of CD4 + splenocytes (left panel) and total numbers of CD4 + splenocytes (right panel) of HMPL-004 and MC treated mice. ( C ) Percentage of CD4 + LPMC (left panel) and total numbers of CD4 + LPMC (right panel) of HMPL-004 and MC treated mice. ( D ) Percentage of CD4 + IL-17 + , or CD4 + IFN-γ + PMA + Ionomycin stimulated splenocytes isolated from HMPL-004 and MC treated mice. ( E ) Total numbers of CD4 + IL-17 + , or CD4 + IFN-γ + splenocytes of HMPL-004 and MC treated mice. Statistical significance was determined by Wilcoxon rank sum test. *, p
    Figure Legend Snippet: HMPL-004 inhibits the early proliferation of CD4 + T cells, and T H 1 and T H 17 polarization in vivo ( A ) Total numbers of mononuclear cells in LP, MLN, and spleen of HMPL-004 and MC treated mice two weeks after T cell transfer (n=11/group for spleens, n=6/group for LPMC, n=8/group for MLN). ( B ) Percentage of CD4 + splenocytes (left panel) and total numbers of CD4 + splenocytes (right panel) of HMPL-004 and MC treated mice. ( C ) Percentage of CD4 + LPMC (left panel) and total numbers of CD4 + LPMC (right panel) of HMPL-004 and MC treated mice. ( D ) Percentage of CD4 + IL-17 + , or CD4 + IFN-γ + PMA + Ionomycin stimulated splenocytes isolated from HMPL-004 and MC treated mice. ( E ) Total numbers of CD4 + IL-17 + , or CD4 + IFN-γ + splenocytes of HMPL-004 and MC treated mice. Statistical significance was determined by Wilcoxon rank sum test. *, p

    Techniques Used: In Vivo, Mouse Assay, Isolation

    7) Product Images from "The disease stage-associated imbalance of Th1/Th2 and Th17/Treg in uterine cervical cancer patients and their recovery with the reduction of tumor burden"

    Article Title: The disease stage-associated imbalance of Th1/Th2 and Th17/Treg in uterine cervical cancer patients and their recovery with the reduction of tumor burden

    Journal: BMC Women's Health

    doi: 10.1186/s12905-020-00972-0

    Th1, Th2, Th17 and Treg cells before and after operation in UCC patients. All the cells were stained with PE- conjugated anti-γ-IFN, anti-IL-4, anti-IL17, anti-FoxP3+ and anti-CD4-FITC. Stained cells were analyzed by flow cytometry analysis using a FACS-CAN cytometer equipped with Cell Quest software. The proportions of each cell type in representative UCC patients are annotated in the figure. The proportions before the operations were a , b , c , and d . The proportions when tested six months after the operation were e , f , g , and h . Th1 cells: a , e ; Th2 cells: b , f ; Th17 cells: c , g ; Treg cells: d , h . The differences of Th1, Th2, Th17 and Treg cell proportions before and after the operations were all significant ( P
    Figure Legend Snippet: Th1, Th2, Th17 and Treg cells before and after operation in UCC patients. All the cells were stained with PE- conjugated anti-γ-IFN, anti-IL-4, anti-IL17, anti-FoxP3+ and anti-CD4-FITC. Stained cells were analyzed by flow cytometry analysis using a FACS-CAN cytometer equipped with Cell Quest software. The proportions of each cell type in representative UCC patients are annotated in the figure. The proportions before the operations were a , b , c , and d . The proportions when tested six months after the operation were e , f , g , and h . Th1 cells: a , e ; Th2 cells: b , f ; Th17 cells: c , g ; Treg cells: d , h . The differences of Th1, Th2, Th17 and Treg cell proportions before and after the operations were all significant ( P

    Techniques Used: Staining, Flow Cytometry, FACS, Cytometry, Software

    Related Articles

    Staining:

    Article Title: Identification and Evaluation of Novel Protective Antigens for the Development of a Candidate Tuberculosis Subunit Vaccine
    Article Snippet: Cells were stained for 10 min with LIVE/DEAD fixable dead cell stain (Invitrogen, UK), followed by surface staining with anti-CD45R/B220, TCRαβ, TCRγδ, CD4, and CD8 (eBioscience). .. Following permeabilization using Cytofix/Cytoperm (BD Biosciences), the cells were stained intracellularly with anti-IFN-γ, anti-TNF-α, anti-IL-2, and anti-IL-17 (eBioscience). .. I-A(b) PPE151–15 phycoerythrin was provided by the NIH Tetramer Facility (Atlanta, GA, USA), and H-2Db PPE15192–200 -allophycocyanin was purchased from ProImmune (Oxford, UK).

    Article Title: Combined Antitumor Effects of Sorafenib and GPC3-CAR T Cells in Mouse Models of Hepatocellular Carcinoma
    Article Snippet: To assess CAR expression, T cells were stained with a goat anti-human biotin-conjugated anti-Fab antibody (Jackson ImmunoResearch), followed by PE-conjugated streptavidin (eBioscience). .. For detection of CD107a and IFN-γ, T cells were fixed, permeabilized, and stained with antibodies against CD107a and IFN-γ (eBioscience). .. To analyze the in vitro proliferation of CAR T cells, a CellTrace Violet cell proliferation kit (Thermo Fisher Scientific) was used for labeling of cells to trace multiple generations via dye dilution by flow cytometry.

    Purification:

    Article Title: Role of TLR2-dependent IL-10 production in the inhibition of the initial IFN-? T cell response to Porphyromonas gingivalis
    Article Snippet: .. Fluorescent-labeled antibodies against CD4 (clone RM4-5), CD8α (clone 53-6.7), CD11b (clone M1/70), CD25 (clone PC61.5), CD69 (clone H12F3), T-bet (clone eBio4B10), Foxp3 (clone FJK-16S), IFN-γ (clone XMG1.2), PD-1 (CD279; clone J43), PD-L1 (CD274; clone MIH5), and purified mouse α-IL-10 (clone JES2A5) were purchased from eBioscience (San Diego, CA, USA). .. Fluorescent-labeled antibodies against Thy1.1 (CD90.1; clone OX-7) and purified mouse α-IL-10R (CD210; clone 1B1.3a) were purchased from BD Biosciences (San Jose, CA, USA).

    Mouse Assay:

    Article Title: Adiponectin deficiency exacerbates lipopolysaccharide/D-galactosamine-induced liver injury in mice
    Article Snippet: The Quantitect PCR probe kit and Quantitect gene assay kit for mice TNF-α, IL-10, and IFN-γ were purchased from Qiagen and used in a real-time PCR analysis of the mouse samples. .. Primers for rat glyceraldehydes-3-phosphate dehydrogenase (GAPDH), TNF-α, IL-10, IFN-γ, and mice GAPDH were designed using the computer program Primer Express (Applied Biosystems, Foster city, CA). .. Dynamo SYBR Green qPCR kit (Finzymes, Espoo, Finland) was used for the real-time PCR analysis of rat TNF-α, IL-10, IFN-γ, GAPDH and mice GAPDH.

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    Thermo Fisher ifn γ
    Plasma concentrations of TNF-α, IL-10 and <t>IFN-γ</t> in mice after GalN/LPS administration. (mean ± SE, n = 6). a P
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    Plasma concentrations of TNF-α, IL-10 and IFN-γ in mice after GalN/LPS administration. (mean ± SE, n = 6). a P

    Journal:

    Article Title: Adiponectin deficiency exacerbates lipopolysaccharide/D-galactosamine-induced liver injury in mice

    doi: 10.3748/wjg.v12.i21.3352

    Figure Lengend Snippet: Plasma concentrations of TNF-α, IL-10 and IFN-γ in mice after GalN/LPS administration. (mean ± SE, n = 6). a P

    Article Snippet: Primers for rat glyceraldehydes-3-phosphate dehydrogenase (GAPDH), TNF-α, IL-10, IFN-γ, and mice GAPDH were designed using the computer program Primer Express (Applied Biosystems, Foster city, CA).

    Techniques: Mouse Assay

    TNF-α, IL-10 and IFN-γ in the liver (A-C) of mice following GalN/LPS administration. (mean ± SE, n = 6). a P

    Journal:

    Article Title: Adiponectin deficiency exacerbates lipopolysaccharide/D-galactosamine-induced liver injury in mice

    doi: 10.3748/wjg.v12.i21.3352

    Figure Lengend Snippet: TNF-α, IL-10 and IFN-γ in the liver (A-C) of mice following GalN/LPS administration. (mean ± SE, n = 6). a P

    Article Snippet: Primers for rat glyceraldehydes-3-phosphate dehydrogenase (GAPDH), TNF-α, IL-10, IFN-γ, and mice GAPDH were designed using the computer program Primer Express (Applied Biosystems, Foster city, CA).

    Techniques: Mouse Assay

    The expressions of adipoR1 and AdipoR2 in Kupffer cells (A). Effect of adiponectin on TNF-α, IL-10 and IFN-γ production by LPS-stimulated Kupffer cells in vitro (B-D). Pre-treated with or without adiponectin (10 mg/L) for 24 h followed

    Journal:

    Article Title: Adiponectin deficiency exacerbates lipopolysaccharide/D-galactosamine-induced liver injury in mice

    doi: 10.3748/wjg.v12.i21.3352

    Figure Lengend Snippet: The expressions of adipoR1 and AdipoR2 in Kupffer cells (A). Effect of adiponectin on TNF-α, IL-10 and IFN-γ production by LPS-stimulated Kupffer cells in vitro (B-D). Pre-treated with or without adiponectin (10 mg/L) for 24 h followed

    Article Snippet: Primers for rat glyceraldehydes-3-phosphate dehydrogenase (GAPDH), TNF-α, IL-10, IFN-γ, and mice GAPDH were designed using the computer program Primer Express (Applied Biosystems, Foster city, CA).

    Techniques: In Vitro

    Il17a −/− Il17f −/− dKO mice exhibit reduced T H 2 cell responses against intranasal allergens. A, Representative FACS plot of CD4 + T cells from the lung. FSC , Forward scatter. B and C, Percentages (Fig 1, B ) and absolute numbers (Fig 1, C ) of CD4 + T-cell subsets. D, Levels of IFN-γ, IL-4, and IL-5 in BAL fluid. The graph shows means ± SEMs. * P

    Journal: The Journal of allergy and clinical immunology

    Article Title: Concomitant suppression of TH2 and TH17 cell responses in allergic asthma by targeting retinoic acid receptor–related orphan receptor γt

    doi: 10.1016/j.jaci.2017.07.050

    Figure Lengend Snippet: Il17a −/− Il17f −/− dKO mice exhibit reduced T H 2 cell responses against intranasal allergens. A, Representative FACS plot of CD4 + T cells from the lung. FSC , Forward scatter. B and C, Percentages (Fig 1, B ) and absolute numbers (Fig 1, C ) of CD4 + T-cell subsets. D, Levels of IFN-γ, IL-4, and IL-5 in BAL fluid. The graph shows means ± SEMs. * P

    Article Snippet: The following antibodies were used for flow cytometric analysis or cell sorting: Alexa Fluor 488–conjugated antibodies to mouse CD62L (MEL-14) and IFN-γ (XMG1.2); fluorescein isothiocyanate–conjugated antibody to human CD45RA (HI100); phycoerythrin-conjugated rat IgG1 (eBRG1; eBioscience); antibodies to mouse CD25 (PC61), IL-17A (TC11-18H10.1), and mouse/human BCL6 (IG191E/A8); peridinin-chlorophyll-protein complex/Cy5.5–conjugated antibodies to mouse CD4 (GK1.5), CD45.1 (A20), and IFN-γ; phycoerythrin/Cy7-conjugated antibodies to mouse IL-13 (eBio13A; eBioscience), mouse/human CD44 (IM7), human CD4 (OKT4), and CD45RO (UCHL1); allophycocyanin (APC)–conjugated antibodies to mouse/human IL-5 (TRFK5), human CD25 (BC96), and IL-4 (8D4-8; eBioscience); Alexa Fluor 647–conjugated antibody to mouse IL-4 (11B11); APC/Cy7-conjugated antibodies to mouse CD4 (GK1.5) and human CD3ε (OKT3); and Pacific blue–conjugated antibodies to mouse CD45.2 (104) and human CD4 (RPA-T4).

    Techniques: Mouse Assay, FACS

    RORγt-deficient mice exhibit reduced T H 2 and T H 17 responses against intranasal allergens. A, Representative FACS plot of lymphocytes from the lung. B and C, Percentages (Fig 2, B ) and absolute numbers (Fig 2, C ) of lung CD4 + T-cell subsets. D, Levels of IFN-γ, IL-4, IL-5, and IL-17A in BAL fluid. E, Absolute numbers of total cells, eosinophils (eo) , macrophages (mac) , lymphocytes (lym) , and neutrophils (neu) in BAL fluid. Data are representative of 3 independent experiments. The graph shows means ± SEMs. * P

    Journal: The Journal of allergy and clinical immunology

    Article Title: Concomitant suppression of TH2 and TH17 cell responses in allergic asthma by targeting retinoic acid receptor–related orphan receptor γt

    doi: 10.1016/j.jaci.2017.07.050

    Figure Lengend Snippet: RORγt-deficient mice exhibit reduced T H 2 and T H 17 responses against intranasal allergens. A, Representative FACS plot of lymphocytes from the lung. B and C, Percentages (Fig 2, B ) and absolute numbers (Fig 2, C ) of lung CD4 + T-cell subsets. D, Levels of IFN-γ, IL-4, IL-5, and IL-17A in BAL fluid. E, Absolute numbers of total cells, eosinophils (eo) , macrophages (mac) , lymphocytes (lym) , and neutrophils (neu) in BAL fluid. Data are representative of 3 independent experiments. The graph shows means ± SEMs. * P

    Article Snippet: The following antibodies were used for flow cytometric analysis or cell sorting: Alexa Fluor 488–conjugated antibodies to mouse CD62L (MEL-14) and IFN-γ (XMG1.2); fluorescein isothiocyanate–conjugated antibody to human CD45RA (HI100); phycoerythrin-conjugated rat IgG1 (eBRG1; eBioscience); antibodies to mouse CD25 (PC61), IL-17A (TC11-18H10.1), and mouse/human BCL6 (IG191E/A8); peridinin-chlorophyll-protein complex/Cy5.5–conjugated antibodies to mouse CD4 (GK1.5), CD45.1 (A20), and IFN-γ; phycoerythrin/Cy7-conjugated antibodies to mouse IL-13 (eBio13A; eBioscience), mouse/human CD44 (IM7), human CD4 (OKT4), and CD45RO (UCHL1); allophycocyanin (APC)–conjugated antibodies to mouse/human IL-5 (TRFK5), human CD25 (BC96), and IL-4 (8D4-8; eBioscience); Alexa Fluor 647–conjugated antibody to mouse IL-4 (11B11); APC/Cy7-conjugated antibodies to mouse CD4 (GK1.5) and human CD3ε (OKT3); and Pacific blue–conjugated antibodies to mouse CD45.2 (104) and human CD4 (RPA-T4).

    Techniques: Mouse Assay, FACS

    Reduction of T H 2 cell responses by UA treatment is IFN-γ independent. A and B, Frequency (Fig 4, A ) and absolute number (Fig 4, B ) of BAL fluid CD4 + T-cell subsets. C, Absolute numbers of total cells, macrophages (mac) , eosinophils (eo) , neutrophils (neu) , and lymphocytes (lym) in BAL fluid. Data are representative of 3 independent experiments. The graph shows means ± SEMs. * P

    Journal: The Journal of allergy and clinical immunology

    Article Title: Concomitant suppression of TH2 and TH17 cell responses in allergic asthma by targeting retinoic acid receptor–related orphan receptor γt

    doi: 10.1016/j.jaci.2017.07.050

    Figure Lengend Snippet: Reduction of T H 2 cell responses by UA treatment is IFN-γ independent. A and B, Frequency (Fig 4, A ) and absolute number (Fig 4, B ) of BAL fluid CD4 + T-cell subsets. C, Absolute numbers of total cells, macrophages (mac) , eosinophils (eo) , neutrophils (neu) , and lymphocytes (lym) in BAL fluid. Data are representative of 3 independent experiments. The graph shows means ± SEMs. * P

    Article Snippet: The following antibodies were used for flow cytometric analysis or cell sorting: Alexa Fluor 488–conjugated antibodies to mouse CD62L (MEL-14) and IFN-γ (XMG1.2); fluorescein isothiocyanate–conjugated antibody to human CD45RA (HI100); phycoerythrin-conjugated rat IgG1 (eBRG1; eBioscience); antibodies to mouse CD25 (PC61), IL-17A (TC11-18H10.1), and mouse/human BCL6 (IG191E/A8); peridinin-chlorophyll-protein complex/Cy5.5–conjugated antibodies to mouse CD4 (GK1.5), CD45.1 (A20), and IFN-γ; phycoerythrin/Cy7-conjugated antibodies to mouse IL-13 (eBio13A; eBioscience), mouse/human CD44 (IM7), human CD4 (OKT4), and CD45RO (UCHL1); allophycocyanin (APC)–conjugated antibodies to mouse/human IL-5 (TRFK5), human CD25 (BC96), and IL-4 (8D4-8; eBioscience); Alexa Fluor 647–conjugated antibody to mouse IL-4 (11B11); APC/Cy7-conjugated antibodies to mouse CD4 (GK1.5) and human CD3ε (OKT3); and Pacific blue–conjugated antibodies to mouse CD45.2 (104) and human CD4 (RPA-T4).

    Techniques:

    IFN-γ and IL-4 production of CD3/CD28 costimulated naive CD4 + T cells from rheumatoid arthritis patients ( n = 9) differentiated during (a) 5-day cultures or (b) 10-day cultures in the presence of IL-7, of IL-4, or of IL-7 and IL-4. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate restimulation for 24 hours. * P

    Journal: Arthritis Research & Therapy

    Article Title: Differentiation of naive CD4+ T cells towards T helper 2 cells is not impaired in rheumatoid arthritis patients

    doi:

    Figure Lengend Snippet: IFN-γ and IL-4 production of CD3/CD28 costimulated naive CD4 + T cells from rheumatoid arthritis patients ( n = 9) differentiated during (a) 5-day cultures or (b) 10-day cultures in the presence of IL-7, of IL-4, or of IL-7 and IL-4. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate restimulation for 24 hours. * P

    Article Snippet: Fluorochrome-labeled anti-IFN-γ, anti-IL-4 and isotype control antibodies (Caltag Laboratories) were added during the permeabilization step (1 μg/ml) to stain the intracellular cytokines.

    Techniques:

    Intracellular detection of IFN-γ and IL-4 by FACS analysis. (a) A representative of seven rheumatoid arthritis (RA) patients of which mean values are shown in (b) . Cytokine production of the CD3/CD28 costimulated CD4 + T cells was assessed after 10 days of culture in the absence or presence of IL-7, of IL-4 or of their combination. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate stimulation for 6 hours in the presence of Brefeldin A. Means ± standard errors of the mean of percentages from cells of RA patients that produce IFN-γ but no IL-4 (Th1) and that produce IL-4 but no IFN-γ (Th2) upon differentiation are shown. * P

    Journal: Arthritis Research & Therapy

    Article Title: Differentiation of naive CD4+ T cells towards T helper 2 cells is not impaired in rheumatoid arthritis patients

    doi:

    Figure Lengend Snippet: Intracellular detection of IFN-γ and IL-4 by FACS analysis. (a) A representative of seven rheumatoid arthritis (RA) patients of which mean values are shown in (b) . Cytokine production of the CD3/CD28 costimulated CD4 + T cells was assessed after 10 days of culture in the absence or presence of IL-7, of IL-4 or of their combination. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate stimulation for 6 hours in the presence of Brefeldin A. Means ± standard errors of the mean of percentages from cells of RA patients that produce IFN-γ but no IL-4 (Th1) and that produce IL-4 but no IFN-γ (Th2) upon differentiation are shown. * P

    Article Snippet: Fluorochrome-labeled anti-IFN-γ, anti-IL-4 and isotype control antibodies (Caltag Laboratories) were added during the permeabilization step (1 μg/ml) to stain the intracellular cytokines.

    Techniques: FACS

    IFN-γ and IL-4 production of CD3/CD28 costimulated naive CD4 + T cells from rheumatoid arthritis patients ( n = 9) stimulated for 5 days in the presence of IL-7. Because of exogenously added IL-4, only the production of IFN-γ is shown for cells cultured in the presence of IL-4 or of IL-7 and IL-4. * P

    Journal: Arthritis Research & Therapy

    Article Title: Differentiation of naive CD4+ T cells towards T helper 2 cells is not impaired in rheumatoid arthritis patients

    doi:

    Figure Lengend Snippet: IFN-γ and IL-4 production of CD3/CD28 costimulated naive CD4 + T cells from rheumatoid arthritis patients ( n = 9) stimulated for 5 days in the presence of IL-7. Because of exogenously added IL-4, only the production of IFN-γ is shown for cells cultured in the presence of IL-4 or of IL-7 and IL-4. * P

    Article Snippet: Fluorochrome-labeled anti-IFN-γ, anti-IL-4 and isotype control antibodies (Caltag Laboratories) were added during the permeabilization step (1 μg/ml) to stain the intracellular cytokines.

    Techniques: Cell Culture

    The production of IFN-γ and IL-4 by CD3/CD28 costimulated naive CD4 + T cells from healthy controls ( n = 9, white bars) and rheumatoid arthritis (RA) patients ( n = 9, black bars) differentiated during 10 days of culture in the presence of IL-7, of IL-4 or of IL-7 and IL-4. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate restimulation for 24 hours. Values are expressed as percentages of cytokine levels produced in the absence of exogenously added cytokines (control values set at 100% for RA patients and controls, respectively, were 47.1 ± 12 ng/ml versus 37.1 ± 5.7 ng/ml for IFN-γ, and 0.89 ± 0.21 ng/ml versus 0.95 ± 0.15 ng/ml for IL-4; not significantly different). # P

    Journal: Arthritis Research & Therapy

    Article Title: Differentiation of naive CD4+ T cells towards T helper 2 cells is not impaired in rheumatoid arthritis patients

    doi:

    Figure Lengend Snippet: The production of IFN-γ and IL-4 by CD3/CD28 costimulated naive CD4 + T cells from healthy controls ( n = 9, white bars) and rheumatoid arthritis (RA) patients ( n = 9, black bars) differentiated during 10 days of culture in the presence of IL-7, of IL-4 or of IL-7 and IL-4. Cytokine production of differentiated cells was analyzed upon ionomycin/phorbol myristate acetate restimulation for 24 hours. Values are expressed as percentages of cytokine levels produced in the absence of exogenously added cytokines (control values set at 100% for RA patients and controls, respectively, were 47.1 ± 12 ng/ml versus 37.1 ± 5.7 ng/ml for IFN-γ, and 0.89 ± 0.21 ng/ml versus 0.95 ± 0.15 ng/ml for IL-4; not significantly different). # P

    Article Snippet: Fluorochrome-labeled anti-IFN-γ, anti-IL-4 and isotype control antibodies (Caltag Laboratories) were added during the permeabilization step (1 μg/ml) to stain the intracellular cytokines.

    Techniques: Produced

    PD-L1 inhibits T-cell secretion of TNF and cytotoxicity. ( a ) MC38-Ova cells were left untreated or treated with IFN γ (5 ng/ml) for 18 h, then flow cytometry was used to analyze expression of MHC-I (H-2Kb) and PD-L1. ( b ) MC38-Ova cells were left untreated or treated with IFN γ (5 ng/ml) for 18 h, then used as targets in a chromium release assay with OT-I T cells (18 h). ( c ) MC38-Ova cells were left untreated or treated with IFN γ (5 ng/ml) for 18 h, then used as targets in a chromium release assay with OT-I T cells (18 h), in the presence of control antibody or anti-PD-1. ( d ) Flow cytometric analysis of MC38-Ova cells transduced with vector, PD-L1, or treated with IFN γ (5 ng/ml for 18 h). ( e ) MC38 PD-L1 cells (1 × 10 5 ) were exposed to Pfn +/+ or Pfn −/− OT-I T cells (1 × 10 4 ), in the presence of control antibody or anti-PD-1 (50 μ g/ml). After 48 h, cytokines in supernatants were measured by CBA. ( f ) MC38 PD-L1 cells (1 × 10 5 ) were exposed to Pfn +/+ or Pfn −/− OT-I T cells (1 × 10 4 ), in the presence of control antibody or anti-PD-1 (50 μ g/ml). After 48 h, survival was analyzed by PI staining and flow cytometry. Error bars represent the mean±S.E.M. of triplicate determinations from a representative experiment, * P

    Journal: Cell Death and Differentiation

    Article Title: PD-L1 and IAPs co-operate to protect tumors from cytotoxic lymphocyte-derived TNF

    doi: 10.1038/cdd.2017.94

    Figure Lengend Snippet: PD-L1 inhibits T-cell secretion of TNF and cytotoxicity. ( a ) MC38-Ova cells were left untreated or treated with IFN γ (5 ng/ml) for 18 h, then flow cytometry was used to analyze expression of MHC-I (H-2Kb) and PD-L1. ( b ) MC38-Ova cells were left untreated or treated with IFN γ (5 ng/ml) for 18 h, then used as targets in a chromium release assay with OT-I T cells (18 h). ( c ) MC38-Ova cells were left untreated or treated with IFN γ (5 ng/ml) for 18 h, then used as targets in a chromium release assay with OT-I T cells (18 h), in the presence of control antibody or anti-PD-1. ( d ) Flow cytometric analysis of MC38-Ova cells transduced with vector, PD-L1, or treated with IFN γ (5 ng/ml for 18 h). ( e ) MC38 PD-L1 cells (1 × 10 5 ) were exposed to Pfn +/+ or Pfn −/− OT-I T cells (1 × 10 4 ), in the presence of control antibody or anti-PD-1 (50 μ g/ml). After 48 h, cytokines in supernatants were measured by CBA. ( f ) MC38 PD-L1 cells (1 × 10 5 ) were exposed to Pfn +/+ or Pfn −/− OT-I T cells (1 × 10 4 ), in the presence of control antibody or anti-PD-1 (50 μ g/ml). After 48 h, survival was analyzed by PI staining and flow cytometry. Error bars represent the mean±S.E.M. of triplicate determinations from a representative experiment, * P

    Article Snippet: Antibodies for immunofluorescence: anti-TNF (Biolegend, clone MP6-XT22), IFN γ (eBiosciences, Scoresby, Victoria, Australia, clone XMG1.2), Tubulin (Rockland).

    Techniques: Flow Cytometry, Cytometry, Expressing, Release Assay, Transduction, Plasmid Preparation, Crocin Bleaching Assay, Staining