gamma interferon ifn γ  (Millipore)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 98
    Name:
    Interferon gamma porcine
    Description:

    Catalog Number:
    I8654
    Price:
    None
    Buy from Supplier


    Structured Review

    Millipore gamma interferon ifn γ
    (A) Quantification of bacterial uptake (%) at 16 h postinfection of murine bone marrow macrophages (BMMΦ) with BCG and M. microti expressing the whole RD1 region (+RD1) or not (−) in the presence or absence of Tween 80. CFU in BMMΦ (B) or after activation with LPS and <t>IFN-γ</t> (C) at 4 h (□) and days 3 ( ) and 6 (▪) after infection with BCG::pYUB412, BCG::RD1, and M. tuberculosis H37Rv. Means and standard deviations are from infections done in quadruplicate wells and are representative of four experiments.

    https://www.bioz.com/result/gamma interferon ifn γ/product/Millipore
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gamma interferon ifn γ - by Bioz Stars, 2021-05
    98/100 stars

    Images

    1) Product Images from "Dissection of ESAT-6 System 1 of Mycobacterium tuberculosis and Impact on Immunogenicity and Virulence "

    Article Title: Dissection of ESAT-6 System 1 of Mycobacterium tuberculosis and Impact on Immunogenicity and Virulence

    Journal: Infection and Immunity

    doi: 10.1128/IAI.74.1.88-98.2006

    (A) Quantification of bacterial uptake (%) at 16 h postinfection of murine bone marrow macrophages (BMMΦ) with BCG and M. microti expressing the whole RD1 region (+RD1) or not (−) in the presence or absence of Tween 80. CFU in BMMΦ (B) or after activation with LPS and IFN-γ (C) at 4 h (□) and days 3 ( ) and 6 (▪) after infection with BCG::pYUB412, BCG::RD1, and M. tuberculosis H37Rv. Means and standard deviations are from infections done in quadruplicate wells and are representative of four experiments.
    Figure Legend Snippet: (A) Quantification of bacterial uptake (%) at 16 h postinfection of murine bone marrow macrophages (BMMΦ) with BCG and M. microti expressing the whole RD1 region (+RD1) or not (−) in the presence or absence of Tween 80. CFU in BMMΦ (B) or after activation with LPS and IFN-γ (C) at 4 h (□) and days 3 ( ) and 6 (▪) after infection with BCG::pYUB412, BCG::RD1, and M. tuberculosis H37Rv. Means and standard deviations are from infections done in quadruplicate wells and are representative of four experiments.

    Techniques Used: Expressing, Activation Assay, Infection

    In vivo bacterial replication of BCG::RD1 in various mouse models: C57BL/6 wild-type mice (open bars), B6;129-LT-α/TNF-α ko mice (light gray bars), inbred 129/Sv IFN-γ receptor ko mice (dark gray bars), and SCID mice (black bars). The ratios of CFU counts at day 28 in spleen (A) and lungs (B) relative to initial dose after intravenous infection with 10 6 CFU are indicated. Each value is the mean of three or four mice, and the error bars represent standard deviations. (C) Histological samples of lungs from C57BL/6 wild type (WT), TNF-α ko, and IFN-γ receptor ko mice, showing compact discrete lesions in lungs of C57BL/6 mice and large diffuse lesions for TNF-α ko mice infected with BCG::RD1 compared to the absence of lesions in wild-type and TNF-α ko mice infected with the BCG vector control. Lungs from IFN-γ receptor ko mice showed lesions with both strains, although bacterial counts were lower for the vector control. Sections were stained with hematoxylin and eosin. Magnification, ×100.
    Figure Legend Snippet: In vivo bacterial replication of BCG::RD1 in various mouse models: C57BL/6 wild-type mice (open bars), B6;129-LT-α/TNF-α ko mice (light gray bars), inbred 129/Sv IFN-γ receptor ko mice (dark gray bars), and SCID mice (black bars). The ratios of CFU counts at day 28 in spleen (A) and lungs (B) relative to initial dose after intravenous infection with 10 6 CFU are indicated. Each value is the mean of three or four mice, and the error bars represent standard deviations. (C) Histological samples of lungs from C57BL/6 wild type (WT), TNF-α ko, and IFN-γ receptor ko mice, showing compact discrete lesions in lungs of C57BL/6 mice and large diffuse lesions for TNF-α ko mice infected with BCG::RD1 compared to the absence of lesions in wild-type and TNF-α ko mice infected with the BCG vector control. Lungs from IFN-γ receptor ko mice showed lesions with both strains, although bacterial counts were lower for the vector control. Sections were stained with hematoxylin and eosin. Magnification, ×100.

    Techniques Used: In Vivo, Mouse Assay, Infection, Plasmid Preparation, Staining

    2) Product Images from "Lentinan Inhibits Tumor Progression by Immunomodulation in a Mouse Model of Bladder Cancer"

    Article Title: Lentinan Inhibits Tumor Progression by Immunomodulation in a Mouse Model of Bladder Cancer

    Journal: Integrative Cancer Therapies

    doi: 10.1177/1534735420946823

    Immunomodulatory effects of LNT on the expression of the cytokines IFN-γ (A), IL-2 (B), TGF-β (C), and IL-10 (D). Data are shown as means ± SD. * P
    Figure Legend Snippet: Immunomodulatory effects of LNT on the expression of the cytokines IFN-γ (A), IL-2 (B), TGF-β (C), and IL-10 (D). Data are shown as means ± SD. * P

    Techniques Used: Expressing

    3) Product Images from "Interferon-? Activates Nuclear Factor-? B in Oligodendrocytes through a Process Mediated by the Unfolded Protein Response"

    Article Title: Interferon-? Activates Nuclear Factor-? B in Oligodendrocytes through a Process Mediated by the Unfolded Protein Response

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0036408

    GADD34 inactivation increased IFN-γ-induced NF-κB activation in oligodendrocytes in IFN-γ-expressing transgenic mice. A, B. CNP and active p65 double immunostaining showed that the immunoreactivity of active p65 was undetectable in oligodendrocytes in the corpus callosum in control IFNγ ^ GADD34 WT mice and IFNγ ^ GADD34 mutant mice. The immunoreactivity of active p65 became detectable in a number of oligodendrocytes in the corpus callosum of IFNγ+GADD34 WT mice, and the number of active p65 positive oligodendrocytes was further increased in the corpus callosum of IFNγ+GADD34 mutant mice. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p
    Figure Legend Snippet: GADD34 inactivation increased IFN-γ-induced NF-κB activation in oligodendrocytes in IFN-γ-expressing transgenic mice. A, B. CNP and active p65 double immunostaining showed that the immunoreactivity of active p65 was undetectable in oligodendrocytes in the corpus callosum in control IFNγ ^ GADD34 WT mice and IFNγ ^ GADD34 mutant mice. The immunoreactivity of active p65 became detectable in a number of oligodendrocytes in the corpus callosum of IFNγ+GADD34 WT mice, and the number of active p65 positive oligodendrocytes was further increased in the corpus callosum of IFNγ+GADD34 mutant mice. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Techniques Used: Activation Assay, Expressing, Transgenic Assay, Mouse Assay, Double Immunostaining, Mutagenesis, Standard Deviation

    Enforced expression of PERKΔC made Oli-neu cells susceptible to the cytotoxicity of IFN-γ. A, B. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Active caspase-3 and DAPI double labeling showed that IFN-γ treatment did not cause Oli-neu cell apoptosis, but significantly increased the number of apoptotic cells in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p
    Figure Legend Snippet: Enforced expression of PERKΔC made Oli-neu cells susceptible to the cytotoxicity of IFN-γ. A, B. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Active caspase-3 and DAPI double labeling showed that IFN-γ treatment did not cause Oli-neu cell apoptosis, but significantly increased the number of apoptotic cells in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Techniques Used: Expressing, Labeling, Standard Deviation

    Enforced expression of PERKΔC impaired IFN-γ-induced NF-κB activation. A, B. The cells were treated with 100 U/ml IFN-γ for 16 hrs. p65 and DAPI double labeling and confocal imaging analysis showed that enforced expression of PERKΔC diminished IFN-γ-induced p65 nucleus translocation in PERKΔC 1 and 10 cells. C, D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. EMSA analysis showed that enforced expression of PERKΔC blocked IFN-γ-induced increase in NF-κB DNA-binding activity in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p
    Figure Legend Snippet: Enforced expression of PERKΔC impaired IFN-γ-induced NF-κB activation. A, B. The cells were treated with 100 U/ml IFN-γ for 16 hrs. p65 and DAPI double labeling and confocal imaging analysis showed that enforced expression of PERKΔC diminished IFN-γ-induced p65 nucleus translocation in PERKΔC 1 and 10 cells. C, D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. EMSA analysis showed that enforced expression of PERKΔC blocked IFN-γ-induced increase in NF-κB DNA-binding activity in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Techniques Used: Expressing, Activation Assay, Labeling, Imaging, Translocation Assay, Binding Assay, Activity Assay, Standard Deviation

    Enforced expression of IκBαΔN blocked IFN-γ-induced NF-κB activation. A. The cells were treated with 100 U/ml IFN-γ for 16 hrs. p65 and DAPI double labeling and confocal imaging analysis showed that p65 remained in the cytoplasm in the untreated Oli-neu cells and that IFN-γ treatment induced translocation of p65 from the cytoplasm to the nucleus in Oli-neu cells. Interestingly, IFN-γ treatment did not result in p65 nucleus translocation in IκBαΔN 1 and 4 cells. B. Quantitative analysis showed that the percentage of Oli-neu cells with p65 nucleus translocation was significantly increased after 16 hrs of 100 U/ml IFN-γ treatment. Nevertheless, enforced expression of IκBαΔN blocked IFN-γ-induced p65 nucleus translocation in IκBαΔN 1 and 4 cells. C, D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. EMSA analysis showed a significant increase in NF-κB DNA-binding activity in Oli-neu cells treated with IFN-γ. Nevertheless, enforced expression of IκBαΔN blocked IFN-γ-induced increase in NF-κB DNA-binding activity in IκBαΔN 1 and 4 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p
    Figure Legend Snippet: Enforced expression of IκBαΔN blocked IFN-γ-induced NF-κB activation. A. The cells were treated with 100 U/ml IFN-γ for 16 hrs. p65 and DAPI double labeling and confocal imaging analysis showed that p65 remained in the cytoplasm in the untreated Oli-neu cells and that IFN-γ treatment induced translocation of p65 from the cytoplasm to the nucleus in Oli-neu cells. Interestingly, IFN-γ treatment did not result in p65 nucleus translocation in IκBαΔN 1 and 4 cells. B. Quantitative analysis showed that the percentage of Oli-neu cells with p65 nucleus translocation was significantly increased after 16 hrs of 100 U/ml IFN-γ treatment. Nevertheless, enforced expression of IκBαΔN blocked IFN-γ-induced p65 nucleus translocation in IκBαΔN 1 and 4 cells. C, D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. EMSA analysis showed a significant increase in NF-κB DNA-binding activity in Oli-neu cells treated with IFN-γ. Nevertheless, enforced expression of IκBαΔN blocked IFN-γ-induced increase in NF-κB DNA-binding activity in IκBαΔN 1 and 4 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Techniques Used: Expressing, Activation Assay, Labeling, Imaging, Translocation Assay, Binding Assay, Activity Assay, Standard Deviation

    Enforced expression of PERKΔC diminished IFN-γ-induced activation of PERK signaling. A. Oli-neu cells were treated with 100 U/ml IFN-γ for 24 hrs. Real-time PCR analysis showed that IFN-γ treatment significantly increased the expression of CHOP and BIP in the cells. B. The cells were treated with 100 U/ml IFN-γ for 8 hrs, 16 hrs, or 24 hrs. Western blot analysis showed that the levels of p-PERK, p-eIF2α, and ATF4 in Oli-neu cells treated with 100 U/ml IFN-γ for 16 hrs were elevated compared to the untreated cells. Moreover, IFN-γ reduced the level of IκBα in Oli-neu cells after 16 hrs of treatment. C. Oli-neu cells were transfected with pBabe-PERKΔC vector encoding Myc epitope-tagged PERKΔC. Immunoblotting for Myc showed that stably transfected cell lines expressed various levels of PERKΔC. PERKΔC 1 and 10 cells expressed high level of PERKΔC. D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. Western blot analysis showed that enforced expression of PERKΔC blocked IFN-γ-induced eIF2α phosphorylation and ATF4 upregulation in PERKΔC 1 and 10 cells. Western blot analysis also showed that enforced expression of PERKΔC diminished IFN-γ-induced reduction of IκBα level in PERKΔC 1 and 10 cells. E. Densitometry analysis of western blot results showed that IFN-γ treatment significantly increased the level of p-eIF2α in Oli-neu cells, but did not affect p-eIF2α level in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p
    Figure Legend Snippet: Enforced expression of PERKΔC diminished IFN-γ-induced activation of PERK signaling. A. Oli-neu cells were treated with 100 U/ml IFN-γ for 24 hrs. Real-time PCR analysis showed that IFN-γ treatment significantly increased the expression of CHOP and BIP in the cells. B. The cells were treated with 100 U/ml IFN-γ for 8 hrs, 16 hrs, or 24 hrs. Western blot analysis showed that the levels of p-PERK, p-eIF2α, and ATF4 in Oli-neu cells treated with 100 U/ml IFN-γ for 16 hrs were elevated compared to the untreated cells. Moreover, IFN-γ reduced the level of IκBα in Oli-neu cells after 16 hrs of treatment. C. Oli-neu cells were transfected with pBabe-PERKΔC vector encoding Myc epitope-tagged PERKΔC. Immunoblotting for Myc showed that stably transfected cell lines expressed various levels of PERKΔC. PERKΔC 1 and 10 cells expressed high level of PERKΔC. D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. Western blot analysis showed that enforced expression of PERKΔC blocked IFN-γ-induced eIF2α phosphorylation and ATF4 upregulation in PERKΔC 1 and 10 cells. Western blot analysis also showed that enforced expression of PERKΔC diminished IFN-γ-induced reduction of IκBα level in PERKΔC 1 and 10 cells. E. Densitometry analysis of western blot results showed that IFN-γ treatment significantly increased the level of p-eIF2α in Oli-neu cells, but did not affect p-eIF2α level in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Techniques Used: Expressing, Activation Assay, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Plasmid Preparation, Stable Transfection, Standard Deviation

    NF-κB activation promoted Oli-neu cell survival in response to IFN-γ. A. The cells were treated with 50 U/ml, 100 U/ml, or 150 U/ml IFN-γ for 24 hrs. MTT assay showed that IFN-γ treatment reduced the number of Oli-neu cells in a dose-dependent manner and that enforced expression of IκBαΔN further reduced the cell numbers. B. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Caspase-3 activity assay showed that IFN-γ treatment did not alter the activity of caspase-3 in Oli-neu cells. Nevertheless, IFN-γ treatment significantly increased the activity of caspase-3 in IκBαΔN 1 and 4 cells. C , D. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Active caspase-3 immunostaining showed that IFN-γ treatment did not change the percentage of active caspase-3 positive cells in Oli-neu cells. Nevertheless, IFN-γ treatment significantly increased the percentage of active caspase-3 positive cells in IκBαΔN 1 and 4 cells. E. BrdU cell proliferation assay showed that the treatment with 100 U/ml IFN-γ for 24 hrs significantly suppressed Oli-neu cell proliferation. Nevertheless, suppression of the NF-κB pathway did not significantly alter the sensitivity of Oli-neu cells to the antiproliferative effects of IFN-γ. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p
    Figure Legend Snippet: NF-κB activation promoted Oli-neu cell survival in response to IFN-γ. A. The cells were treated with 50 U/ml, 100 U/ml, or 150 U/ml IFN-γ for 24 hrs. MTT assay showed that IFN-γ treatment reduced the number of Oli-neu cells in a dose-dependent manner and that enforced expression of IκBαΔN further reduced the cell numbers. B. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Caspase-3 activity assay showed that IFN-γ treatment did not alter the activity of caspase-3 in Oli-neu cells. Nevertheless, IFN-γ treatment significantly increased the activity of caspase-3 in IκBαΔN 1 and 4 cells. C , D. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Active caspase-3 immunostaining showed that IFN-γ treatment did not change the percentage of active caspase-3 positive cells in Oli-neu cells. Nevertheless, IFN-γ treatment significantly increased the percentage of active caspase-3 positive cells in IκBαΔN 1 and 4 cells. E. BrdU cell proliferation assay showed that the treatment with 100 U/ml IFN-γ for 24 hrs significantly suppressed Oli-neu cell proliferation. Nevertheless, suppression of the NF-κB pathway did not significantly alter the sensitivity of Oli-neu cells to the antiproliferative effects of IFN-γ. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Techniques Used: Activation Assay, MTT Assay, Expressing, Caspase-3 Activity Assay, Activity Assay, Immunostaining, BrdU Cell Proliferation Assay, Standard Deviation

    4) Product Images from "Impairment of Immunoproteasome Function by ?5i/LMP7 Subunit Deficiency Results in Severe Enterovirus Myocarditis"

    Article Title: Impairment of Immunoproteasome Function by ?5i/LMP7 Subunit Deficiency Results in Severe Enterovirus Myocarditis

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1002233

    Accumulation of poly-ub protein conjugates in IP-deficient mice. (A) Primary cardiomyocytes and B-cell depleted splenocytes from β5i/LMP7 +/+ and β5i/LMP7 -/- mice were incubated with IFN-γ for indicated time points. Lack of IP formation resulted in accumulation of poly-ub substrates and of oxidant-damaged proteins (staining of carbonyl groups) upon IFN-γ exposure in both cell populations (cell purity is depicted in Fig. S1 ). GAPDH illustrates equal protein loading. Images are representative from at least two independent experiments. (B) NFκB p50 levels were determined in whole cell homogenates from IFN-γ (100 U/ml for 24 h) treated primary cardiomyocytes and B-cell depleted splenocytes upon 30 min TNF-α exposure. Data are shown as fold increase of p50 levels in comparison to untreated control cells (representative mean from triplicates from at least two independent experiments). (C) Immunoblot of CVB3-infected cardiac homogenates from five individual β5i/LMP7 +/+ and β5i/LMP7 -/- mice (d8 p.i.) stained for poly-ub-conjugates (from the same transfer; representative for n = 10 mice). Densitometry was performed for individual lanes for these five different mice (normalization to GAPDH control) yielding increased levels of poly-ub signals in β5i/LMP7 -/- mice (mean±SEM). (D) Formation of poly-ub containing ALIS (white arrowheads) was visualized by immunofluorescence: heart cryosections were stained with FK1 for poly-ub (green) and Hoechst (blue). Here, slides are representative for n = 6 mice from two independent experiments (more data is illustrated in Fig. S3 ). (E) To localize poly-ub conjugates within the injured myocardium, immunohistology staining of ubiquitin was performed in control and CVB3-infected mice (d8 p.i.). Representative tissue sections are shown. For more information see Fig. S4 to this figure. (F) CVB3-challenged cardiac lysates from from five individual β5i/LMP7 +/+ and β5i/LMP7 -/- mice (d8 p.i.; representative for n = 9 mice) stained for oxidant-damaged proteins by staining of carbonyl groups. Quantitative evaluation of oxy-staining was performed as described above. All statistical data are mean±SEM; * p
    Figure Legend Snippet: Accumulation of poly-ub protein conjugates in IP-deficient mice. (A) Primary cardiomyocytes and B-cell depleted splenocytes from β5i/LMP7 +/+ and β5i/LMP7 -/- mice were incubated with IFN-γ for indicated time points. Lack of IP formation resulted in accumulation of poly-ub substrates and of oxidant-damaged proteins (staining of carbonyl groups) upon IFN-γ exposure in both cell populations (cell purity is depicted in Fig. S1 ). GAPDH illustrates equal protein loading. Images are representative from at least two independent experiments. (B) NFκB p50 levels were determined in whole cell homogenates from IFN-γ (100 U/ml for 24 h) treated primary cardiomyocytes and B-cell depleted splenocytes upon 30 min TNF-α exposure. Data are shown as fold increase of p50 levels in comparison to untreated control cells (representative mean from triplicates from at least two independent experiments). (C) Immunoblot of CVB3-infected cardiac homogenates from five individual β5i/LMP7 +/+ and β5i/LMP7 -/- mice (d8 p.i.) stained for poly-ub-conjugates (from the same transfer; representative for n = 10 mice). Densitometry was performed for individual lanes for these five different mice (normalization to GAPDH control) yielding increased levels of poly-ub signals in β5i/LMP7 -/- mice (mean±SEM). (D) Formation of poly-ub containing ALIS (white arrowheads) was visualized by immunofluorescence: heart cryosections were stained with FK1 for poly-ub (green) and Hoechst (blue). Here, slides are representative for n = 6 mice from two independent experiments (more data is illustrated in Fig. S3 ). (E) To localize poly-ub conjugates within the injured myocardium, immunohistology staining of ubiquitin was performed in control and CVB3-infected mice (d8 p.i.). Representative tissue sections are shown. For more information see Fig. S4 to this figure. (F) CVB3-challenged cardiac lysates from from five individual β5i/LMP7 +/+ and β5i/LMP7 -/- mice (d8 p.i.; representative for n = 9 mice) stained for oxidant-damaged proteins by staining of carbonyl groups. Quantitative evaluation of oxy-staining was performed as described above. All statistical data are mean±SEM; * p

    Techniques Used: Mouse Assay, Incubation, Staining, Infection, Immunofluorescence

    Characterization of 20S proteasomes in IP-deficient mice in CVB3-myocarditis. (A) Quantitative real-time PCR: β1i/LMP2, β2i/MECL-1, and β5i/LMP7 mRNA expression in the hearts of β5i/LMP7 +/+ and β5i/LMP7 -/- mice at d4 and d8 p.i. Data shown are representative means of n = 5 mice±SEM. (B) 20S proteasomes were isolated from cardiac tissue and subjected to SDS-PAGE followed by immunoblotting (proteasome subunit α6 is a loading control). Silver staining of isolated 20S proteasomes illustrates equal loading. Data shown are representative for two different 20S proteasome purifications (n = 10 mice per experiment). (C) Primary embryonic cardiomyocytes were stimulated with 100 U/ml IFN-γ for indicated time points and the expression of 20S proteasome immunosubunits was determined in whole tissue homogenates. β1i/LMP2 antibody shows a cross-reaction to constitutive β1 subunit. GAPDH illustrates equal protein loading.
    Figure Legend Snippet: Characterization of 20S proteasomes in IP-deficient mice in CVB3-myocarditis. (A) Quantitative real-time PCR: β1i/LMP2, β2i/MECL-1, and β5i/LMP7 mRNA expression in the hearts of β5i/LMP7 +/+ and β5i/LMP7 -/- mice at d4 and d8 p.i. Data shown are representative means of n = 5 mice±SEM. (B) 20S proteasomes were isolated from cardiac tissue and subjected to SDS-PAGE followed by immunoblotting (proteasome subunit α6 is a loading control). Silver staining of isolated 20S proteasomes illustrates equal loading. Data shown are representative for two different 20S proteasome purifications (n = 10 mice per experiment). (C) Primary embryonic cardiomyocytes were stimulated with 100 U/ml IFN-γ for indicated time points and the expression of 20S proteasome immunosubunits was determined in whole tissue homogenates. β1i/LMP2 antibody shows a cross-reaction to constitutive β1 subunit. GAPDH illustrates equal protein loading.

    Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction, Expressing, Isolation, SDS Page, Silver Staining

    5) Product Images from "cPLA2 Regulates the Expression of Type I Interferons and Intracellular Immunity to Chlamydia trachomatis *"

    Article Title: cPLA2 Regulates the Expression of Type I Interferons and Intracellular Immunity to Chlamydia trachomatis *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.103010

    cPLA2 −/− MEFs are responsive to exogenous interferon. cPLA2 +/+ and cPLA2 −/− MEFs were treated with 100 units/ml IFN-γ and tested for the expression of IRG proteins ( A ) and the ability contain chlamydial replication,
    Figure Legend Snippet: cPLA2 −/− MEFs are responsive to exogenous interferon. cPLA2 +/+ and cPLA2 −/− MEFs were treated with 100 units/ml IFN-γ and tested for the expression of IRG proteins ( A ) and the ability contain chlamydial replication,

    Techniques Used: Expressing

    6) Product Images from "Potent Inhibition of Jun?n Virus Infection by Interferon in Murine Cells"

    Article Title: Potent Inhibition of Jun?n Virus Infection by Interferon in Murine Cells

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0002933

    Interferon sensitivity of JUNV in different cell lines. (A) mouse embryonic fibroblast cells (MEF), (B) Vero cells (ATCC) and (C) human lung carcinoma epithelial A549 cells (ATCC) were treated with IFNs at the indicated concentrations for 16 h. Vero cells and A549 cells were treated with human IFN-α2b (Schering), IFN-β (PBL) or IFN-γ (Sigma), while MEF cells were treated with mouse IFN-β (PBL), respectively. Cells were then infected with VSV, Candid#1 JUNV or Romero JUNV at an MOI of 0.1 PFU/cell. IFNs were supplemented after virus infection. During Romero and Candid#1 JUNV infection, supernatants were collected at 3 days p.i. and assayed for virus production by plaque assay. During VSV infection, supernatants were collected at 16 h.p.i.. Dotted lines indicate the limitation of plaque assay. Data represent the mean of three experiments ±SEM.
    Figure Legend Snippet: Interferon sensitivity of JUNV in different cell lines. (A) mouse embryonic fibroblast cells (MEF), (B) Vero cells (ATCC) and (C) human lung carcinoma epithelial A549 cells (ATCC) were treated with IFNs at the indicated concentrations for 16 h. Vero cells and A549 cells were treated with human IFN-α2b (Schering), IFN-β (PBL) or IFN-γ (Sigma), while MEF cells were treated with mouse IFN-β (PBL), respectively. Cells were then infected with VSV, Candid#1 JUNV or Romero JUNV at an MOI of 0.1 PFU/cell. IFNs were supplemented after virus infection. During Romero and Candid#1 JUNV infection, supernatants were collected at 3 days p.i. and assayed for virus production by plaque assay. During VSV infection, supernatants were collected at 16 h.p.i.. Dotted lines indicate the limitation of plaque assay. Data represent the mean of three experiments ±SEM.

    Techniques Used: Infection, Plaque Assay

    7) Product Images from "Impaired Proinflammatory Response in Stringently Defined Otitis-prone Children During Viral Upper Respiratory Infections"

    Article Title: Impaired Proinflammatory Response in Stringently Defined Otitis-prone Children During Viral Upper Respiratory Infections

    Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

    doi: 10.1093/cid/ciy750

    Comparison of cytokine and chemokine levels between upper respiratory tract infection (URI) and acute otitis media (AOM) visits for non–otitis-prone (NOP) children. IL-8, IL-6, IL-10, MIP-1α, IFN-γ, TNF-α, IL-1β, MCP-1, RANTES, IL-2, and IL-17 levels in nasal washes were measured for URI visits and the AOM visits following the URIs in 18 NOP children using a Milliplex MAP human cytokine/chemokine magnetic bead panel kit. The cytokine/chemokine concentration (pg/ml) was then normalized by the concentration of the total protein (µg/ml) in the nasal wash, which was quantitated using a micro bicinchoninic acid protein assay kit. A ratio-paired t -test was used to analyze the differences of the normalized cytokine/chemokine levels (pg per µg of total protein). Data are represented as geometric means ± 95% confidence intervals. Abbreviations: AOM, acute otitis media; IFN, interferon; IL, interleukin; MCP, monocyte chemoattractant protein; MIP, macrophage inflammatory protein; NOP, non–otitis-prone; RANTES, regulated on activation, normal T-cell–expressed and –secreted; TNF, tumor necrosis factor; URI, upper respiratory tract infection.
    Figure Legend Snippet: Comparison of cytokine and chemokine levels between upper respiratory tract infection (URI) and acute otitis media (AOM) visits for non–otitis-prone (NOP) children. IL-8, IL-6, IL-10, MIP-1α, IFN-γ, TNF-α, IL-1β, MCP-1, RANTES, IL-2, and IL-17 levels in nasal washes were measured for URI visits and the AOM visits following the URIs in 18 NOP children using a Milliplex MAP human cytokine/chemokine magnetic bead panel kit. The cytokine/chemokine concentration (pg/ml) was then normalized by the concentration of the total protein (µg/ml) in the nasal wash, which was quantitated using a micro bicinchoninic acid protein assay kit. A ratio-paired t -test was used to analyze the differences of the normalized cytokine/chemokine levels (pg per µg of total protein). Data are represented as geometric means ± 95% confidence intervals. Abbreviations: AOM, acute otitis media; IFN, interferon; IL, interleukin; MCP, monocyte chemoattractant protein; MIP, macrophage inflammatory protein; NOP, non–otitis-prone; RANTES, regulated on activation, normal T-cell–expressed and –secreted; TNF, tumor necrosis factor; URI, upper respiratory tract infection.

    Techniques Used: Infection, Concentration Assay, Bicinchoninic Acid Protein Assay, Activation Assay

    Comparison of cytokine and chemokine levels between upper respiratory tract infection (URI) and acute otitis media (AOM) visits for stringently defined otitis-prone (sOP) children. IL-8, IL-6, IL-10, MIP-1α, IFN-γ, TNF-α, IL-1β, MCP-1, RANTES, IL-2, and IL-17 levels in nasal washes were measured for URI visits and the AOM visits following the URIs in 21 sOP children using a Milliplex MAP Human cytokine/chemokine magnetic bead panel kit. The cytokine/chemokine concentration (pg/ml) was then normalized by the concentration of the total protein (µg/ml) in the nasal wash, which was quantitated with a micro bicinchoninic acid protein assay kit. A ratio-paired t -test was used to analyze the differences of the normalized cytokine/chemokine levels (pg per µg of total protein). Data are represented as geometric means ± 95% confidence intervals. Abbreviations: AOM, acute otitis media; IFN, interferon; IL, interleukin; MCP, monocyte chemoattractant protein; MIP, macrophage inflammatory protein; RANTES, regulated on activation, normal T-cell–expressed and –secreted; sOP, stringently-defined otitis-prone; TNF, tumor necrosis factor; URI, upper respiratory tract infection.
    Figure Legend Snippet: Comparison of cytokine and chemokine levels between upper respiratory tract infection (URI) and acute otitis media (AOM) visits for stringently defined otitis-prone (sOP) children. IL-8, IL-6, IL-10, MIP-1α, IFN-γ, TNF-α, IL-1β, MCP-1, RANTES, IL-2, and IL-17 levels in nasal washes were measured for URI visits and the AOM visits following the URIs in 21 sOP children using a Milliplex MAP Human cytokine/chemokine magnetic bead panel kit. The cytokine/chemokine concentration (pg/ml) was then normalized by the concentration of the total protein (µg/ml) in the nasal wash, which was quantitated with a micro bicinchoninic acid protein assay kit. A ratio-paired t -test was used to analyze the differences of the normalized cytokine/chemokine levels (pg per µg of total protein). Data are represented as geometric means ± 95% confidence intervals. Abbreviations: AOM, acute otitis media; IFN, interferon; IL, interleukin; MCP, monocyte chemoattractant protein; MIP, macrophage inflammatory protein; RANTES, regulated on activation, normal T-cell–expressed and –secreted; sOP, stringently-defined otitis-prone; TNF, tumor necrosis factor; URI, upper respiratory tract infection.

    Techniques Used: Infection, Concentration Assay, Bicinchoninic Acid Protein Assay, Activation Assay

    Logistic regression analysis of the relationship between cytokine and chemokine levels and the rate of stringently-defined otitis-prone (sOP) children at upper respiratory tract infection (URI) only visits. IL-8, IL-6, IL-10, MIP-1α, IFN-γ, TNF-α, IL-1β, MCP-1, IFN-α2, RANTES, IL-2, IL-4, and IL-17 levels in nasal washes were measured for 29 URI visits of 18 sOP children and 58 URI visits of 49 non–otitis-prone children using a Milliplex MAP human cytokine/chemokine magnetic bead panel kit. The cytokine/chemokine concentration (pg/ml) was then normalized by the concentration of the total protein (µg/ml) in the nasal wash, which was quantitated with a micro bicinchoninic acid protein assay kit. The cytokine level (pg per µg of total protein) was log-transformed and, where possible, the log mean of the 2 duplicates was used as a single outcome. Cytokine/chemokine levels below the detection limits were treated as missing, because the protein concentrations were generally very low or not detectable, suggesting poor success in obtaining an adequate nasal wash sample. For 10 of 13 cytokine/chemokines, repeated measures were available. Kernel smoothers were used to assess the relationship between cytokine/chemokine levels and sOP classification frequencies. In general, little change in the sOP classification frequencies was evident at lower cytokine/chemokine levels. In some cases, rapid changes in sOP classification frequencies were observed at higher cytokine/chemokine levels. In addition, even after log transformation, outlier cytokine/chemokine levels were evident, with potentially disproportionate influences on any regression models. Therefore, cytokine/chemokine levels were transformed into ranks and then normalized into the unit interval. To model the rapid change in sOP classification frequencies at higher cytokine/chemokine levels, a power transformation was then applied to the rescaled levels. This predictor was then used in a logistic regression model: P (OP ) = ɸ ( η ), where, for transformed cytokine/chemokine level X, η = β 0 + β 1 X k and ɸ ( η ) = e η /(1 + e η ). Application of the Bayesian information criterion yielded an optimal choice of κ near 8 for those cytokines/chemokines with a significant association with the sOP classification frequency, so this parameter was used for the remaining analysis. The data contain subject-level repeated measures, but since sOP is fixed for each subject, logistic regression models that incorporate subject-level correlations, such as generalized estimating equation models, are unable to produce stable model estimates. Therefore, in order to assess significance, a bootstrap procedure with subject-level resampling was used. To model the rapid change in sOP rates at higher cytokine levels, a power transformation was then applied to the rescaled cytokine levels. In order to assess significance, a bootstrap procedure with subject-level resampling was used. Abbreviations: AOM, acute otitis media; IFN, interferon; IL, interleukin; MCP, monocyte chemoattractant protein; MIP, macrophage inflammatory protein; RANTES, regulated on activation, normal T-cell–expressed and –secreted; sOP, stringently-defined otitis-prone; TNF, tumor necrosis factor; URI, upper respiratory tract infection.
    Figure Legend Snippet: Logistic regression analysis of the relationship between cytokine and chemokine levels and the rate of stringently-defined otitis-prone (sOP) children at upper respiratory tract infection (URI) only visits. IL-8, IL-6, IL-10, MIP-1α, IFN-γ, TNF-α, IL-1β, MCP-1, IFN-α2, RANTES, IL-2, IL-4, and IL-17 levels in nasal washes were measured for 29 URI visits of 18 sOP children and 58 URI visits of 49 non–otitis-prone children using a Milliplex MAP human cytokine/chemokine magnetic bead panel kit. The cytokine/chemokine concentration (pg/ml) was then normalized by the concentration of the total protein (µg/ml) in the nasal wash, which was quantitated with a micro bicinchoninic acid protein assay kit. The cytokine level (pg per µg of total protein) was log-transformed and, where possible, the log mean of the 2 duplicates was used as a single outcome. Cytokine/chemokine levels below the detection limits were treated as missing, because the protein concentrations were generally very low or not detectable, suggesting poor success in obtaining an adequate nasal wash sample. For 10 of 13 cytokine/chemokines, repeated measures were available. Kernel smoothers were used to assess the relationship between cytokine/chemokine levels and sOP classification frequencies. In general, little change in the sOP classification frequencies was evident at lower cytokine/chemokine levels. In some cases, rapid changes in sOP classification frequencies were observed at higher cytokine/chemokine levels. In addition, even after log transformation, outlier cytokine/chemokine levels were evident, with potentially disproportionate influences on any regression models. Therefore, cytokine/chemokine levels were transformed into ranks and then normalized into the unit interval. To model the rapid change in sOP classification frequencies at higher cytokine/chemokine levels, a power transformation was then applied to the rescaled levels. This predictor was then used in a logistic regression model: P (OP ) = ɸ ( η ), where, for transformed cytokine/chemokine level X, η = β 0 + β 1 X k and ɸ ( η ) = e η /(1 + e η ). Application of the Bayesian information criterion yielded an optimal choice of κ near 8 for those cytokines/chemokines with a significant association with the sOP classification frequency, so this parameter was used for the remaining analysis. The data contain subject-level repeated measures, but since sOP is fixed for each subject, logistic regression models that incorporate subject-level correlations, such as generalized estimating equation models, are unable to produce stable model estimates. Therefore, in order to assess significance, a bootstrap procedure with subject-level resampling was used. To model the rapid change in sOP rates at higher cytokine levels, a power transformation was then applied to the rescaled cytokine levels. In order to assess significance, a bootstrap procedure with subject-level resampling was used. Abbreviations: AOM, acute otitis media; IFN, interferon; IL, interleukin; MCP, monocyte chemoattractant protein; MIP, macrophage inflammatory protein; RANTES, regulated on activation, normal T-cell–expressed and –secreted; sOP, stringently-defined otitis-prone; TNF, tumor necrosis factor; URI, upper respiratory tract infection.

    Techniques Used: Infection, Concentration Assay, Bicinchoninic Acid Protein Assay, Transformation Assay, Activation Assay

    8) Product Images from "Human Papillomavirus 16 E5 Modulates the Expression of Host MicroRNAs"

    Article Title: Human Papillomavirus 16 E5 Modulates the Expression of Host MicroRNAs

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0021646

    Effect of miRNA transfections on p63 expression and activation of TNF-α or IFN-γ signaling. HaCaT-E5 and –pMSG cells were transfected with 20 nM pre-miR-203 or scrambled miRNA negative control (scr neg.ctrl). After overnight incubation, the cells were serum-starved for 24 h, and subsequently treated with 1 µM dexamethasone to induce E5 expression. Forty-eight hours after induction the cells were harvested and the cell lysates analysed for p63 expression by western blotting. Equal loading was confirmed by probing the same filter with β-actin. The numbers below each lane represent p63 protein expression fold change normalized to β-actin relative to scr neg.ctrl of pMSG cells ( A ). HaCaT-E5 and -pMSG cells were transfected with scr neg.ctrl miRNA, and with either pre-miR-203 ( B ) or with anti-miR-146a ( C ). The transfection procedure was as described for A . Before harvesting, the cells were treated with IFN-γ (10 ng/ml) ( B ) or TNF-α (20 ng/ml) ( C ) for indicated periods of time. The cell lysates were analyzed with western blotting for phospho-p38 (p–p38), phospho-STAT1 (p-STAT1; B ), or phospho-ERK1/2 (p-ERK1/2; C ). The levels of total p38, STAT1, ERK1/2, and β-actin were determined as controls.
    Figure Legend Snippet: Effect of miRNA transfections on p63 expression and activation of TNF-α or IFN-γ signaling. HaCaT-E5 and –pMSG cells were transfected with 20 nM pre-miR-203 or scrambled miRNA negative control (scr neg.ctrl). After overnight incubation, the cells were serum-starved for 24 h, and subsequently treated with 1 µM dexamethasone to induce E5 expression. Forty-eight hours after induction the cells were harvested and the cell lysates analysed for p63 expression by western blotting. Equal loading was confirmed by probing the same filter with β-actin. The numbers below each lane represent p63 protein expression fold change normalized to β-actin relative to scr neg.ctrl of pMSG cells ( A ). HaCaT-E5 and -pMSG cells were transfected with scr neg.ctrl miRNA, and with either pre-miR-203 ( B ) or with anti-miR-146a ( C ). The transfection procedure was as described for A . Before harvesting, the cells were treated with IFN-γ (10 ng/ml) ( B ) or TNF-α (20 ng/ml) ( C ) for indicated periods of time. The cell lysates were analyzed with western blotting for phospho-p38 (p–p38), phospho-STAT1 (p-STAT1; B ), or phospho-ERK1/2 (p-ERK1/2; C ). The levels of total p38, STAT1, ERK1/2, and β-actin were determined as controls.

    Techniques Used: Transfection, Expressing, Activation Assay, Negative Control, Incubation, Western Blot

    9) Product Images from "DNA Sensing-Independent Inhibition of Herpes Simplex Virus 1 Replication by DAI/ZBP1"

    Article Title: DNA Sensing-Independent Inhibition of Herpes Simplex Virus 1 Replication by DAI/ZBP1

    Journal: Journal of Virology

    doi: 10.1128/JVI.02860-12

    DAI knockdown enhances HSV-1 replication in HepG2 cells. (A) HepG2 cells were untreated or treated with IFN-γ (100 ng/ml) or mock-infected or infected with HSV-1 at an MOI of 3 for indicated times. The levels of DAI and β-actin mRNAs were
    Figure Legend Snippet: DAI knockdown enhances HSV-1 replication in HepG2 cells. (A) HepG2 cells were untreated or treated with IFN-γ (100 ng/ml) or mock-infected or infected with HSV-1 at an MOI of 3 for indicated times. The levels of DAI and β-actin mRNAs were

    Techniques Used: Infection

    10) Product Images from "Immunity-related GTPase M (IRGM) Proteins Influence the Localization of Guanylate-binding Protein 2 (GBP2) by Modulating Macroautophagy"

    Article Title: Immunity-related GTPase M (IRGM) Proteins Influence the Localization of Guanylate-binding Protein 2 (GBP2) by Modulating Macroautophagy

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M111.251967

    Altered localization of Gbp2 in cells deficient for Irgm1 or Irgm3. A , 3T3 MEFs or primary bone marrow macrophages of the indicated genotypes were activated with 100 units/ml IFN-γ for 24 h and then processed for immunostaining with anti-Gbp2
    Figure Legend Snippet: Altered localization of Gbp2 in cells deficient for Irgm1 or Irgm3. A , 3T3 MEFs or primary bone marrow macrophages of the indicated genotypes were activated with 100 units/ml IFN-γ for 24 h and then processed for immunostaining with anti-Gbp2

    Techniques Used: Immunostaining

    Irgm1 deficiency leads to impaired autophagic flux. A , MEFs of the indicated genotypes were or were not activated with IFN-γ for 24 h and then processed for Western blotting with anti-p62 antibodies. Shown is one representative of four experiments.
    Figure Legend Snippet: Irgm1 deficiency leads to impaired autophagic flux. A , MEFs of the indicated genotypes were or were not activated with IFN-γ for 24 h and then processed for Western blotting with anti-p62 antibodies. Shown is one representative of four experiments.

    Techniques Used: Western Blot

    IRGM deficiency leads to colocalization of Gbp2 with ubiquitin and p62. MEFs of the indicated genotypes were activated with IFN-γ for 24 h and then processed for immunostaining with anti-p62 antibody ( A ) or FK2 (anti-ubiquitin) antibody ( B ). Shown
    Figure Legend Snippet: IRGM deficiency leads to colocalization of Gbp2 with ubiquitin and p62. MEFs of the indicated genotypes were activated with IFN-γ for 24 h and then processed for immunostaining with anti-p62 antibody ( A ) or FK2 (anti-ubiquitin) antibody ( B ). Shown

    Techniques Used: Immunostaining

    Gbp2 colocalizes with GKS IRG proteins in IRGM-deficient cells. 3T3 MEFs of the indicated genotypes were activated with 100 units/ml IFN-γ for 24 h and then processed for immunostaining with anti-Gbp2 and Irga6 antibodies ( A ) or anti-Irgb6 and
    Figure Legend Snippet: Gbp2 colocalizes with GKS IRG proteins in IRGM-deficient cells. 3T3 MEFs of the indicated genotypes were activated with 100 units/ml IFN-γ for 24 h and then processed for immunostaining with anti-Gbp2 and Irga6 antibodies ( A ) or anti-Irgb6 and

    Techniques Used: Immunostaining

    Gbp2 localizes to autophagosomal structures independent of IRGM activity. A , poly-Lys-63-ubiquitinated proteins were precipitated from IFN-γ treated MEF lysates of the indicated genotypes using TUBE 1. Proteins were separated by SDS-PAGE and blotted
    Figure Legend Snippet: Gbp2 localizes to autophagosomal structures independent of IRGM activity. A , poly-Lys-63-ubiquitinated proteins were precipitated from IFN-γ treated MEF lysates of the indicated genotypes using TUBE 1. Proteins were separated by SDS-PAGE and blotted

    Techniques Used: Activity Assay, SDS Page

    IRGM deficiency leads to increased levels of Lys-63-ubiquitinated proteins, including Irga6. Poly-Lys-63-ubiquitinated proteins were precipitated from IFN-γ-treated MEF lysates of the indicated genotypes using TUBE 1-conjugated resin. Proteins
    Figure Legend Snippet: IRGM deficiency leads to increased levels of Lys-63-ubiquitinated proteins, including Irga6. Poly-Lys-63-ubiquitinated proteins were precipitated from IFN-γ-treated MEF lysates of the indicated genotypes using TUBE 1-conjugated resin. Proteins

    Techniques Used:

    IRGM deficiency leads to an increase in Gbp2-tagged autophagosome accumulation. A , MEFs of the indicated genotypes were activated with IFN-γ for 24 h and then processed for immunostaining with anti-LC3 antibodies. Shown are representative two-dimensional
    Figure Legend Snippet: IRGM deficiency leads to an increase in Gbp2-tagged autophagosome accumulation. A , MEFs of the indicated genotypes were activated with IFN-γ for 24 h and then processed for immunostaining with anti-LC3 antibodies. Shown are representative two-dimensional

    Techniques Used: Immunostaining

    11) Product Images from "MAP kinase phosphatase 1 controls innate immune responses and suppresses endotoxic shock"

    Article Title: MAP kinase phosphatase 1 controls innate immune responses and suppresses endotoxic shock

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20051794

    IFN-γ enhances activation of p38 and JNK by inhibiting MKP-1 expression. Resident peritoneal macrophages were treated with or without 50 U/ml IFN-γ overnight before LPS stimulation (100 ng/ml) at the indicated times. The levels of MKP-1 protein and phosphorylated MAP kinases were examined using Western blotting. Please note that the order for the + IFN-γ samples is from right to left.
    Figure Legend Snippet: IFN-γ enhances activation of p38 and JNK by inhibiting MKP-1 expression. Resident peritoneal macrophages were treated with or without 50 U/ml IFN-γ overnight before LPS stimulation (100 ng/ml) at the indicated times. The levels of MKP-1 protein and phosphorylated MAP kinases were examined using Western blotting. Please note that the order for the + IFN-γ samples is from right to left.

    Techniques Used: Activation Assay, Expressing, Western Blot

    Knockout of Mkp-1 alters cytokine expression in macrophages. (A) Cytokine production by IFN-γ–primed resident peritoneal macrophages. Resident peritoneal macrophages primed with IFN-γ overnight were stimulated with LPS for 4 and 6 h. Cytokine concentrations in the medium were analyzed by ELISA. Data are presented as the mean ± SEM ( n = 3 in each group). *, Mkp-1 −/− different from Mkp-1 +/+ , P
    Figure Legend Snippet: Knockout of Mkp-1 alters cytokine expression in macrophages. (A) Cytokine production by IFN-γ–primed resident peritoneal macrophages. Resident peritoneal macrophages primed with IFN-γ overnight were stimulated with LPS for 4 and 6 h. Cytokine concentrations in the medium were analyzed by ELISA. Data are presented as the mean ± SEM ( n = 3 in each group). *, Mkp-1 −/− different from Mkp-1 +/+ , P

    Techniques Used: Knock-Out, Expressing, Enzyme-linked Immunosorbent Assay

    Knockout of Mkp-1 shifts the pattern of cytokine expression in splenocytes and dendritic cells. (A) Cytokine expression in LPS-stimulated splenocytes. Splenocytes isolated from Mkp-1 +/+ and Mkp-1 −/− mice were stimulated with 100 ng/ml LPS for the indicated times, and cytokine concentrations in the medium were assayed by ELISA. IFN-γ levels were measured 24 h after LPS treatment. Data are presented as the mean ± SEM ( n = 3–9 for each group). *, Mkp-1 −/− different from Mkp-1 +/+ , P
    Figure Legend Snippet: Knockout of Mkp-1 shifts the pattern of cytokine expression in splenocytes and dendritic cells. (A) Cytokine expression in LPS-stimulated splenocytes. Splenocytes isolated from Mkp-1 +/+ and Mkp-1 −/− mice were stimulated with 100 ng/ml LPS for the indicated times, and cytokine concentrations in the medium were assayed by ELISA. IFN-γ levels were measured 24 h after LPS treatment. Data are presented as the mean ± SEM ( n = 3–9 for each group). *, Mkp-1 −/− different from Mkp-1 +/+ , P

    Techniques Used: Knock-Out, Expressing, Isolation, Mouse Assay, Enzyme-linked Immunosorbent Assay

    Related Articles

    Activation Assay:

    Article Title: Microglia Induce Neurotoxicity via Intraneuronal Zn2+ Release and a K+ Current Surge
    Article Snippet: For the Zn2+ imaging and electrophysiological studies, trans-well inserts containing microglia were placed on top of neuron-containing coverslips in 24-well plates immediately prior to microglial activation. .. To ensure a robust and consistent activation of microglia, 10 U/mL interferon-γ (IFN-γ, Chemicon, Temecula, CA) and 1 μg/mL lipopolysaccharide were added directly into the well inserts ( ). .. Chemical activation of this microglial cell line leads to activation of both iNOS and NADPH oxidase in less than 30–60 min ( ; ).

    Article Title: Impaired Proinflammatory Response in Stringently Defined Otitis-prone Children During Viral Upper Respiratory Infections
    Article Snippet: Viral ribonucleic acid was isolated using extraction kits (Qiagen) and amplified by real-time reverse transcription polymerase chain reaction using RSVa, RSVb, PIV3, influenza A, influenza B, adenovirus b, adenovirus c, enterovirus, and rhinovirus kits specific for each virus (Primer Design, United Kingdom), as previously described [ ]. .. Interleukin (IL)-8; IL-6; IL-10; macrophage inflammatory protein (MIP)-1α; interferon (IFN)-γ; tumor necrosis factor (TNF)-α; IL-1β; monocyte chemoattractant protein (MCP)-1; IFN-α2; regulated on activation, normal T-cell–expressed and –secreted (RANTES); IL-2; IL-4; and IL-17 levels in nasal washes were detected using the Milliplex MAP human cytokine/chemokine magnetic bead panel kit (EMD Millipore, Billerica, Massachusetts). .. The plates were read on a Luminex 200 and a High-Throughput Fluidics instrument (Bio-Rad, Hercules, California).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Lentinan Inhibits Tumor Progression by Immunomodulation in a Mouse Model of Bladder Cancer
    Article Snippet: A total of 10,000 events acquired in the gate were analyzed by FlowJo v7.6.2 software (Tree Star, San Carlos, CA, USA). .. Enzyme-Linked Immunosorbent Assay (ELISA)The expression levels of transforming growth factor (TGF)-β, interferon (IFN)-γ, interleukin (IL)-2, and IL-10 in the lysate of the single-cell suspension of splenocytes and tumor tissues were measured by ELISA kits (Sigma-Aldrich Co., St. Louis, MO, USA). ..

    Expressing:

    Article Title: Lentinan Inhibits Tumor Progression by Immunomodulation in a Mouse Model of Bladder Cancer
    Article Snippet: A total of 10,000 events acquired in the gate were analyzed by FlowJo v7.6.2 software (Tree Star, San Carlos, CA, USA). .. Enzyme-Linked Immunosorbent Assay (ELISA)The expression levels of transforming growth factor (TGF)-β, interferon (IFN)-γ, interleukin (IL)-2, and IL-10 in the lysate of the single-cell suspension of splenocytes and tumor tissues were measured by ELISA kits (Sigma-Aldrich Co., St. Louis, MO, USA). ..

    Incubation:

    Article Title: Dissection of ESAT-6 System 1 of Mycobacterium tuberculosis and Impact on Immunogenicity and Virulence
    Article Snippet: .. Activated macrophages were prepared by incubation with 100 U of gamma interferon (IFN-γ) and 10 ng of lipopolysaccharide (LPS) (Sigma, Saint Louis, MO)/ml 24 h prior to infection. .. Alveolar epithelial A549 cells (American Type Culture Collection) were cultured in RPMI 1640 supplemented with 10% heat-inactivated fetal calf serum.

    Infection:

    Article Title: Dissection of ESAT-6 System 1 of Mycobacterium tuberculosis and Impact on Immunogenicity and Virulence
    Article Snippet: .. Activated macrophages were prepared by incubation with 100 U of gamma interferon (IFN-γ) and 10 ng of lipopolysaccharide (LPS) (Sigma, Saint Louis, MO)/ml 24 h prior to infection. .. Alveolar epithelial A549 cells (American Type Culture Collection) were cultured in RPMI 1640 supplemented with 10% heat-inactivated fetal calf serum.

    Article Title: Identification of Francisella tularensis Live Vaccine Strain CuZn Superoxide Dismutase as Critical for Resistance to Extracellularly Generated Reactive Oxygen Species
    Article Snippet: .. A murine alveolar macrophage cell line, MH-S (ATTC CRL-2019), was left untreated or treated with 50 or 100 ng/ml gamma interferon (IFN-γ) (Sigma Aldrich, St. Louis, MO) for 16 h before and after infection at a multiplicity of infection (MOI) of 100 with F . tularensis LVS, sod mutants, or the transcomplemented strain. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 98
    Millipore ifn γ
    GADD34 inactivation increased <t>IFN-γ-induced</t> NF-κB activation in oligodendrocytes in IFN-γ-expressing transgenic mice. A, B. CNP and active p65 double immunostaining showed that the immunoreactivity of active p65 was undetectable in oligodendrocytes in the corpus callosum in control IFNγ ^ GADD34 WT mice and IFNγ ^ GADD34 mutant mice. The immunoreactivity of active p65 became detectable in a number of oligodendrocytes in the corpus callosum of IFNγ+GADD34 WT mice, and the number of active p65 positive oligodendrocytes was further increased in the corpus callosum of IFNγ+GADD34 mutant mice. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p
    Ifn γ, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ifn γ/product/Millipore
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ifn γ - by Bioz Stars, 2021-05
    98/100 stars
      Buy from Supplier

    99
    Millipore hiv ifn γ elispot 96 well polyvinylidene difluoride backed elispot plates
    Validation of the qPCR assay using whole blood. qPCR for MIG and IP10 can detect CMV (a) MTB (b) and <t>HIV</t> (C) -specific immune responses using only 25 (for CMV) and 50 uL (for HIV/MTB) of whole blood. 100, 25 and 10 uL of whole blood from healthy donors with was diluted 1∶5 with R10 media and stimulated with 10 ng/mL <t>IFN-γ</t> or ten-fold dilutions of CMV whole cell lysate (10 uL/ml)(Sigma) for 16 hours (a). 50 and 25 ul of whole blood diluted 1∶5 with R10 media were stimulated with 1, 5, 10, or 20 µl of 0.16 mg/ml ESAT-6 and CFP-10 peptide pools for 16 hours (b). Representative example of 50 µl of whole blood taken from a HIV positive patient without symptoms suggestive of active TB and an RD1 positive <t>Elispot,</t> diluted 1∶5 with R10 media and stimulated with ESAT-6 and CFP-10 (both at final concentrations of 8 ug/ml) and HIV peptide pools (at a final concentration of 10 ug/ml).
    Hiv Ifn γ Elispot 96 Well Polyvinylidene Difluoride Backed Elispot Plates, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hiv ifn γ elispot 96 well polyvinylidene difluoride backed elispot plates/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hiv ifn γ elispot 96 well polyvinylidene difluoride backed elispot plates - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

    N/A
    This ELISA antibody pair detects human IFN gamma R1 Other species not yet determined
      Buy from Supplier

    Image Search Results


    GADD34 inactivation increased IFN-γ-induced NF-κB activation in oligodendrocytes in IFN-γ-expressing transgenic mice. A, B. CNP and active p65 double immunostaining showed that the immunoreactivity of active p65 was undetectable in oligodendrocytes in the corpus callosum in control IFNγ ^ GADD34 WT mice and IFNγ ^ GADD34 mutant mice. The immunoreactivity of active p65 became detectable in a number of oligodendrocytes in the corpus callosum of IFNγ+GADD34 WT mice, and the number of active p65 positive oligodendrocytes was further increased in the corpus callosum of IFNγ+GADD34 mutant mice. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Journal: PLoS ONE

    Article Title: Interferon-? Activates Nuclear Factor-? B in Oligodendrocytes through a Process Mediated by the Unfolded Protein Response

    doi: 10.1371/journal.pone.0036408

    Figure Lengend Snippet: GADD34 inactivation increased IFN-γ-induced NF-κB activation in oligodendrocytes in IFN-γ-expressing transgenic mice. A, B. CNP and active p65 double immunostaining showed that the immunoreactivity of active p65 was undetectable in oligodendrocytes in the corpus callosum in control IFNγ ^ GADD34 WT mice and IFNγ ^ GADD34 mutant mice. The immunoreactivity of active p65 became detectable in a number of oligodendrocytes in the corpus callosum of IFNγ+GADD34 WT mice, and the number of active p65 positive oligodendrocytes was further increased in the corpus callosum of IFNγ+GADD34 mutant mice. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Article Snippet: NF-κB Electrophoretic Mobility Shift Assay (EMSA) Oli-neu cells treated with IFN-γ (EMD Biosciences) for 16 hrs were rinsed with ice-cold PBS.

    Techniques: Activation Assay, Expressing, Transgenic Assay, Mouse Assay, Double Immunostaining, Mutagenesis, Standard Deviation

    Enforced expression of PERKΔC made Oli-neu cells susceptible to the cytotoxicity of IFN-γ. A, B. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Active caspase-3 and DAPI double labeling showed that IFN-γ treatment did not cause Oli-neu cell apoptosis, but significantly increased the number of apoptotic cells in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Journal: PLoS ONE

    Article Title: Interferon-? Activates Nuclear Factor-? B in Oligodendrocytes through a Process Mediated by the Unfolded Protein Response

    doi: 10.1371/journal.pone.0036408

    Figure Lengend Snippet: Enforced expression of PERKΔC made Oli-neu cells susceptible to the cytotoxicity of IFN-γ. A, B. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Active caspase-3 and DAPI double labeling showed that IFN-γ treatment did not cause Oli-neu cell apoptosis, but significantly increased the number of apoptotic cells in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Article Snippet: NF-κB Electrophoretic Mobility Shift Assay (EMSA) Oli-neu cells treated with IFN-γ (EMD Biosciences) for 16 hrs were rinsed with ice-cold PBS.

    Techniques: Expressing, Labeling, Standard Deviation

    Enforced expression of PERKΔC impaired IFN-γ-induced NF-κB activation. A, B. The cells were treated with 100 U/ml IFN-γ for 16 hrs. p65 and DAPI double labeling and confocal imaging analysis showed that enforced expression of PERKΔC diminished IFN-γ-induced p65 nucleus translocation in PERKΔC 1 and 10 cells. C, D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. EMSA analysis showed that enforced expression of PERKΔC blocked IFN-γ-induced increase in NF-κB DNA-binding activity in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Journal: PLoS ONE

    Article Title: Interferon-? Activates Nuclear Factor-? B in Oligodendrocytes through a Process Mediated by the Unfolded Protein Response

    doi: 10.1371/journal.pone.0036408

    Figure Lengend Snippet: Enforced expression of PERKΔC impaired IFN-γ-induced NF-κB activation. A, B. The cells were treated with 100 U/ml IFN-γ for 16 hrs. p65 and DAPI double labeling and confocal imaging analysis showed that enforced expression of PERKΔC diminished IFN-γ-induced p65 nucleus translocation in PERKΔC 1 and 10 cells. C, D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. EMSA analysis showed that enforced expression of PERKΔC blocked IFN-γ-induced increase in NF-κB DNA-binding activity in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Article Snippet: NF-κB Electrophoretic Mobility Shift Assay (EMSA) Oli-neu cells treated with IFN-γ (EMD Biosciences) for 16 hrs were rinsed with ice-cold PBS.

    Techniques: Expressing, Activation Assay, Labeling, Imaging, Translocation Assay, Binding Assay, Activity Assay, Standard Deviation

    Enforced expression of IκBαΔN blocked IFN-γ-induced NF-κB activation. A. The cells were treated with 100 U/ml IFN-γ for 16 hrs. p65 and DAPI double labeling and confocal imaging analysis showed that p65 remained in the cytoplasm in the untreated Oli-neu cells and that IFN-γ treatment induced translocation of p65 from the cytoplasm to the nucleus in Oli-neu cells. Interestingly, IFN-γ treatment did not result in p65 nucleus translocation in IκBαΔN 1 and 4 cells. B. Quantitative analysis showed that the percentage of Oli-neu cells with p65 nucleus translocation was significantly increased after 16 hrs of 100 U/ml IFN-γ treatment. Nevertheless, enforced expression of IκBαΔN blocked IFN-γ-induced p65 nucleus translocation in IκBαΔN 1 and 4 cells. C, D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. EMSA analysis showed a significant increase in NF-κB DNA-binding activity in Oli-neu cells treated with IFN-γ. Nevertheless, enforced expression of IκBαΔN blocked IFN-γ-induced increase in NF-κB DNA-binding activity in IκBαΔN 1 and 4 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Journal: PLoS ONE

    Article Title: Interferon-? Activates Nuclear Factor-? B in Oligodendrocytes through a Process Mediated by the Unfolded Protein Response

    doi: 10.1371/journal.pone.0036408

    Figure Lengend Snippet: Enforced expression of IκBαΔN blocked IFN-γ-induced NF-κB activation. A. The cells were treated with 100 U/ml IFN-γ for 16 hrs. p65 and DAPI double labeling and confocal imaging analysis showed that p65 remained in the cytoplasm in the untreated Oli-neu cells and that IFN-γ treatment induced translocation of p65 from the cytoplasm to the nucleus in Oli-neu cells. Interestingly, IFN-γ treatment did not result in p65 nucleus translocation in IκBαΔN 1 and 4 cells. B. Quantitative analysis showed that the percentage of Oli-neu cells with p65 nucleus translocation was significantly increased after 16 hrs of 100 U/ml IFN-γ treatment. Nevertheless, enforced expression of IκBαΔN blocked IFN-γ-induced p65 nucleus translocation in IκBαΔN 1 and 4 cells. C, D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. EMSA analysis showed a significant increase in NF-κB DNA-binding activity in Oli-neu cells treated with IFN-γ. Nevertheless, enforced expression of IκBαΔN blocked IFN-γ-induced increase in NF-κB DNA-binding activity in IκBαΔN 1 and 4 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Article Snippet: NF-κB Electrophoretic Mobility Shift Assay (EMSA) Oli-neu cells treated with IFN-γ (EMD Biosciences) for 16 hrs were rinsed with ice-cold PBS.

    Techniques: Expressing, Activation Assay, Labeling, Imaging, Translocation Assay, Binding Assay, Activity Assay, Standard Deviation

    Enforced expression of PERKΔC diminished IFN-γ-induced activation of PERK signaling. A. Oli-neu cells were treated with 100 U/ml IFN-γ for 24 hrs. Real-time PCR analysis showed that IFN-γ treatment significantly increased the expression of CHOP and BIP in the cells. B. The cells were treated with 100 U/ml IFN-γ for 8 hrs, 16 hrs, or 24 hrs. Western blot analysis showed that the levels of p-PERK, p-eIF2α, and ATF4 in Oli-neu cells treated with 100 U/ml IFN-γ for 16 hrs were elevated compared to the untreated cells. Moreover, IFN-γ reduced the level of IκBα in Oli-neu cells after 16 hrs of treatment. C. Oli-neu cells were transfected with pBabe-PERKΔC vector encoding Myc epitope-tagged PERKΔC. Immunoblotting for Myc showed that stably transfected cell lines expressed various levels of PERKΔC. PERKΔC 1 and 10 cells expressed high level of PERKΔC. D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. Western blot analysis showed that enforced expression of PERKΔC blocked IFN-γ-induced eIF2α phosphorylation and ATF4 upregulation in PERKΔC 1 and 10 cells. Western blot analysis also showed that enforced expression of PERKΔC diminished IFN-γ-induced reduction of IκBα level in PERKΔC 1 and 10 cells. E. Densitometry analysis of western blot results showed that IFN-γ treatment significantly increased the level of p-eIF2α in Oli-neu cells, but did not affect p-eIF2α level in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Journal: PLoS ONE

    Article Title: Interferon-? Activates Nuclear Factor-? B in Oligodendrocytes through a Process Mediated by the Unfolded Protein Response

    doi: 10.1371/journal.pone.0036408

    Figure Lengend Snippet: Enforced expression of PERKΔC diminished IFN-γ-induced activation of PERK signaling. A. Oli-neu cells were treated with 100 U/ml IFN-γ for 24 hrs. Real-time PCR analysis showed that IFN-γ treatment significantly increased the expression of CHOP and BIP in the cells. B. The cells were treated with 100 U/ml IFN-γ for 8 hrs, 16 hrs, or 24 hrs. Western blot analysis showed that the levels of p-PERK, p-eIF2α, and ATF4 in Oli-neu cells treated with 100 U/ml IFN-γ for 16 hrs were elevated compared to the untreated cells. Moreover, IFN-γ reduced the level of IκBα in Oli-neu cells after 16 hrs of treatment. C. Oli-neu cells were transfected with pBabe-PERKΔC vector encoding Myc epitope-tagged PERKΔC. Immunoblotting for Myc showed that stably transfected cell lines expressed various levels of PERKΔC. PERKΔC 1 and 10 cells expressed high level of PERKΔC. D. The cells were treated with 100 U/ml IFN-γ for 16 hrs. Western blot analysis showed that enforced expression of PERKΔC blocked IFN-γ-induced eIF2α phosphorylation and ATF4 upregulation in PERKΔC 1 and 10 cells. Western blot analysis also showed that enforced expression of PERKΔC diminished IFN-γ-induced reduction of IκBα level in PERKΔC 1 and 10 cells. E. Densitometry analysis of western blot results showed that IFN-γ treatment significantly increased the level of p-eIF2α in Oli-neu cells, but did not affect p-eIF2α level in PERKΔC 1 and 10 cells. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Article Snippet: NF-κB Electrophoretic Mobility Shift Assay (EMSA) Oli-neu cells treated with IFN-γ (EMD Biosciences) for 16 hrs were rinsed with ice-cold PBS.

    Techniques: Expressing, Activation Assay, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Plasmid Preparation, Stable Transfection, Standard Deviation

    NF-κB activation promoted Oli-neu cell survival in response to IFN-γ. A. The cells were treated with 50 U/ml, 100 U/ml, or 150 U/ml IFN-γ for 24 hrs. MTT assay showed that IFN-γ treatment reduced the number of Oli-neu cells in a dose-dependent manner and that enforced expression of IκBαΔN further reduced the cell numbers. B. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Caspase-3 activity assay showed that IFN-γ treatment did not alter the activity of caspase-3 in Oli-neu cells. Nevertheless, IFN-γ treatment significantly increased the activity of caspase-3 in IκBαΔN 1 and 4 cells. C , D. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Active caspase-3 immunostaining showed that IFN-γ treatment did not change the percentage of active caspase-3 positive cells in Oli-neu cells. Nevertheless, IFN-γ treatment significantly increased the percentage of active caspase-3 positive cells in IκBαΔN 1 and 4 cells. E. BrdU cell proliferation assay showed that the treatment with 100 U/ml IFN-γ for 24 hrs significantly suppressed Oli-neu cell proliferation. Nevertheless, suppression of the NF-κB pathway did not significantly alter the sensitivity of Oli-neu cells to the antiproliferative effects of IFN-γ. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Journal: PLoS ONE

    Article Title: Interferon-? Activates Nuclear Factor-? B in Oligodendrocytes through a Process Mediated by the Unfolded Protein Response

    doi: 10.1371/journal.pone.0036408

    Figure Lengend Snippet: NF-κB activation promoted Oli-neu cell survival in response to IFN-γ. A. The cells were treated with 50 U/ml, 100 U/ml, or 150 U/ml IFN-γ for 24 hrs. MTT assay showed that IFN-γ treatment reduced the number of Oli-neu cells in a dose-dependent manner and that enforced expression of IκBαΔN further reduced the cell numbers. B. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Caspase-3 activity assay showed that IFN-γ treatment did not alter the activity of caspase-3 in Oli-neu cells. Nevertheless, IFN-γ treatment significantly increased the activity of caspase-3 in IκBαΔN 1 and 4 cells. C , D. The cells were treated with 100 U/ml IFN-γ for 24 hrs. Active caspase-3 immunostaining showed that IFN-γ treatment did not change the percentage of active caspase-3 positive cells in Oli-neu cells. Nevertheless, IFN-γ treatment significantly increased the percentage of active caspase-3 positive cells in IκBαΔN 1 and 4 cells. E. BrdU cell proliferation assay showed that the treatment with 100 U/ml IFN-γ for 24 hrs significantly suppressed Oli-neu cell proliferation. Nevertheless, suppression of the NF-κB pathway did not significantly alter the sensitivity of Oli-neu cells to the antiproliferative effects of IFN-γ. The experiments were repeated at least three times, error bars represent standard deviation, asterisk p

    Article Snippet: NF-κB Electrophoretic Mobility Shift Assay (EMSA) Oli-neu cells treated with IFN-γ (EMD Biosciences) for 16 hrs were rinsed with ice-cold PBS.

    Techniques: Activation Assay, MTT Assay, Expressing, Caspase-3 Activity Assay, Activity Assay, Immunostaining, BrdU Cell Proliferation Assay, Standard Deviation

    Validation of the qPCR assay using whole blood. qPCR for MIG and IP10 can detect CMV (a) MTB (b) and HIV (C) -specific immune responses using only 25 (for CMV) and 50 uL (for HIV/MTB) of whole blood. 100, 25 and 10 uL of whole blood from healthy donors with was diluted 1∶5 with R10 media and stimulated with 10 ng/mL IFN-γ or ten-fold dilutions of CMV whole cell lysate (10 uL/ml)(Sigma) for 16 hours (a). 50 and 25 ul of whole blood diluted 1∶5 with R10 media were stimulated with 1, 5, 10, or 20 µl of 0.16 mg/ml ESAT-6 and CFP-10 peptide pools for 16 hours (b). Representative example of 50 µl of whole blood taken from a HIV positive patient without symptoms suggestive of active TB and an RD1 positive Elispot, diluted 1∶5 with R10 media and stimulated with ESAT-6 and CFP-10 (both at final concentrations of 8 ug/ml) and HIV peptide pools (at a final concentration of 10 ug/ml).

    Journal: PLoS ONE

    Article Title: A Molecular Assay for Sensitive Detection of Pathogen-Specific T-Cells

    doi: 10.1371/journal.pone.0020606

    Figure Lengend Snippet: Validation of the qPCR assay using whole blood. qPCR for MIG and IP10 can detect CMV (a) MTB (b) and HIV (C) -specific immune responses using only 25 (for CMV) and 50 uL (for HIV/MTB) of whole blood. 100, 25 and 10 uL of whole blood from healthy donors with was diluted 1∶5 with R10 media and stimulated with 10 ng/mL IFN-γ or ten-fold dilutions of CMV whole cell lysate (10 uL/ml)(Sigma) for 16 hours (a). 50 and 25 ul of whole blood diluted 1∶5 with R10 media were stimulated with 1, 5, 10, or 20 µl of 0.16 mg/ml ESAT-6 and CFP-10 peptide pools for 16 hours (b). Representative example of 50 µl of whole blood taken from a HIV positive patient without symptoms suggestive of active TB and an RD1 positive Elispot, diluted 1∶5 with R10 media and stimulated with ESAT-6 and CFP-10 (both at final concentrations of 8 ug/ml) and HIV peptide pools (at a final concentration of 10 ug/ml).

    Article Snippet: CMV IFN-γ Elispot, MTB Region of Difference-1 (RD1) IFN-γ Elispot and HIV IFN-γ Elispot 96-well polyvinylidene difluoride-backed Elispot plates (MAIP S45, Millipore) were coated overnight at 4°C with 100 µl anti-IFN-γ antibody (1-D1k, 0.5 µg/ml, MabTech, Sweden).

    Techniques: Real-time Polymerase Chain Reaction, Enzyme-linked Immunospot, Concentration Assay