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  • 90
    ProSci Incorporated α bim
    α Bim, supplied by ProSci Incorporated, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/α bim/product/ProSci Incorporated
    Average 90 stars, based on 1 article reviews
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    96
    JASCO Inc jasco hplc
    Jasco Hplc, supplied by JASCO Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/jasco hplc/product/JASCO Inc
    Average 96 stars, based on 1 article reviews
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    92
    R&D Systems mouse s100a9
    U937 cells ( A ), J774A.1 cells ( B ), primary bone marrow derived macrophages or BMDM ( C ) and primary mouse alveolar macrophages ( D ) were infected with IAV. U937 cells were infected at 1 MOI, whereas J774A.1, BMDM and primary alvelolar macrophages were infected at 2 MOI. At indicated post-infection time-periods the medium supernatant was collected to assess levels of <t>S100A9</t> protein by ELISA. The values shown represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test.
    Mouse S100a9, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse s100a9/product/R&D Systems
    Average 92 stars, based on 1 article reviews
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    mouse s100a9 - by Bioz Stars, 2024-06
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    93
    Cell Signaling Technology Inc 2065s
    U937 cells ( A ), J774A.1 cells ( B ), primary bone marrow derived macrophages or BMDM ( C ) and primary mouse alveolar macrophages ( D ) were infected with IAV. U937 cells were infected at 1 MOI, whereas J774A.1, BMDM and primary alvelolar macrophages were infected at 2 MOI. At indicated post-infection time-periods the medium supernatant was collected to assess levels of <t>S100A9</t> protein by ELISA. The values shown represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test.
    2065s, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/2065s/product/Cell Signaling Technology Inc
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    89
    Bio-Techne corporation s100a9
    U937 cells ( A ), J774A.1 cells ( B ), primary bone marrow derived macrophages or BMDM ( C ) and primary mouse alveolar macrophages ( D ) were infected with IAV. U937 cells were infected at 1 MOI, whereas J774A.1, BMDM and primary alvelolar macrophages were infected at 2 MOI. At indicated post-infection time-periods the medium supernatant was collected to assess levels of <t>S100A9</t> protein by ELISA. The values shown represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test.
    S100a9, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s100a9/product/Bio-Techne corporation
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    s100a9 - by Bioz Stars, 2024-06
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    Image Search Results


    U937 cells ( A ), J774A.1 cells ( B ), primary bone marrow derived macrophages or BMDM ( C ) and primary mouse alveolar macrophages ( D ) were infected with IAV. U937 cells were infected at 1 MOI, whereas J774A.1, BMDM and primary alvelolar macrophages were infected at 2 MOI. At indicated post-infection time-periods the medium supernatant was collected to assess levels of S100A9 protein by ELISA. The values shown represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test.

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: U937 cells ( A ), J774A.1 cells ( B ), primary bone marrow derived macrophages or BMDM ( C ) and primary mouse alveolar macrophages ( D ) were infected with IAV. U937 cells were infected at 1 MOI, whereas J774A.1, BMDM and primary alvelolar macrophages were infected at 2 MOI. At indicated post-infection time-periods the medium supernatant was collected to assess levels of S100A9 protein by ELISA. The values shown represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test.

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay, Standard Deviation

    ( A ) Primary bone marrow derived macrophages or BMDMs isolated from wild-type (WT), TLR2 knockout (KO), TLR4 KO and TRIF KO mice were infected with IAV (2 MOI). At 24 h post-infection time-period the medium supernatant was collected to assess levels of S100A9 protein by ELISA. ( B ) RT-PCR analysis of S100A9 expression in IAV infected WT and TRIF KO BMDMs. ( C ) BMDMs isolated from WT and TLR3 KO mice were infected with IAV (2 MOI). At indicated post-infection time-periods the medium supernatant was collected to assess levels of S100A9 protein by ELISA. ( D ) Mouse alveolar macrophage cell-line MH-S was transfected with either control siRNA or DDX21 specific siRNA. At 48 h post-transfection, cells were infected with IAV (2 MOI). At indicated post-infection time-period RT-PCR analysis was performed to examine expression of DDX21 in IAV infected control and DDX21 silenced cells. ( E ) MH-S cells transfected with either control siRNA or DDX21 specific siRNA were infected with IAV (2 MOI). At indicated post-infection time-period the medium supernatant was collected to assess levels of S100A9 protein by ELISA. ( F ) BMDMs isolated from WT and TLR7 KO mice were infected with IAV (2 MOI). At indicated post-infection time-periods the medium supernatant was collected to assess levels of S100A9 protein by ELISA. The values shown in (A), (C), (E) and (F) represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test. Each RT-PCR data (B and D) is a representative of three independent experiments with similar results.

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: ( A ) Primary bone marrow derived macrophages or BMDMs isolated from wild-type (WT), TLR2 knockout (KO), TLR4 KO and TRIF KO mice were infected with IAV (2 MOI). At 24 h post-infection time-period the medium supernatant was collected to assess levels of S100A9 protein by ELISA. ( B ) RT-PCR analysis of S100A9 expression in IAV infected WT and TRIF KO BMDMs. ( C ) BMDMs isolated from WT and TLR3 KO mice were infected with IAV (2 MOI). At indicated post-infection time-periods the medium supernatant was collected to assess levels of S100A9 protein by ELISA. ( D ) Mouse alveolar macrophage cell-line MH-S was transfected with either control siRNA or DDX21 specific siRNA. At 48 h post-transfection, cells were infected with IAV (2 MOI). At indicated post-infection time-period RT-PCR analysis was performed to examine expression of DDX21 in IAV infected control and DDX21 silenced cells. ( E ) MH-S cells transfected with either control siRNA or DDX21 specific siRNA were infected with IAV (2 MOI). At indicated post-infection time-period the medium supernatant was collected to assess levels of S100A9 protein by ELISA. ( F ) BMDMs isolated from WT and TLR7 KO mice were infected with IAV (2 MOI). At indicated post-infection time-periods the medium supernatant was collected to assess levels of S100A9 protein by ELISA. The values shown in (A), (C), (E) and (F) represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test. Each RT-PCR data (B and D) is a representative of three independent experiments with similar results.

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Derivative Assay, Isolation, Knock-Out, Infection, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Transfection, Standard Deviation

    Human U937 macrophages were incubated with purified recombinant human S100A9 protein (10 µg/ml) for 6 h and 12 h. The medium supernatant was collected to assess levels of human TNF-α (TNF) ( A ) and human IL-6 ( B ) by ELISA. Mouse J774A.1 macrophages were incubated with purified recombinant mouse S100A9 protein (5 µg/ml) for 6 h and 12 h. The medium supernatant was collected to assess levels of mouse TNF ( C ) and mouse IL-6 ( D ) by ELISA. The values represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test. Vehicle control cells (veh) were incubated with HBSS buffer.

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: Human U937 macrophages were incubated with purified recombinant human S100A9 protein (10 µg/ml) for 6 h and 12 h. The medium supernatant was collected to assess levels of human TNF-α (TNF) ( A ) and human IL-6 ( B ) by ELISA. Mouse J774A.1 macrophages were incubated with purified recombinant mouse S100A9 protein (5 µg/ml) for 6 h and 12 h. The medium supernatant was collected to assess levels of mouse TNF ( C ) and mouse IL-6 ( D ) by ELISA. The values represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test. Vehicle control cells (veh) were incubated with HBSS buffer.

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Incubation, Purification, Recombinant, Enzyme-linked Immunosorbent Assay, Standard Deviation

    (A) Mouse J774A.1 macrophages were infected with IAV (2 MOI) in the presence of either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab). At indicated post-infection time-periods the medium supernatant was collected to assess levels of mouse IL-6 by ELISA. ( B ) Primary bone marrow derived macrophages (BMDM) isolated from wild-type (WT) and S100A9 knockout (KO) mice were infected with IAV (2 MOI). The medium supernatant was collected to assess levels of mouse IL-6 by ELISA. ( C ) WT and S100A9 KO BMDM were infected with IAV (2 MOI). At the indicated post-infection time-period, medium supernatant was collected to assess levels of mouse TNF-α(TNF) by ELISA. ( D ) S100A9 KO BMDMs were infected with IAV (2 MOI) in the presence of purified recombinant mouse S100A9 protein (5 µg/ml). Medium supernatant was collected from infected cells to assess levels of mouse TNF and IL-6 by ELISA. Vehicle control cells (veh) were incubated with HBSS buffer. The values represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test.

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: (A) Mouse J774A.1 macrophages were infected with IAV (2 MOI) in the presence of either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab). At indicated post-infection time-periods the medium supernatant was collected to assess levels of mouse IL-6 by ELISA. ( B ) Primary bone marrow derived macrophages (BMDM) isolated from wild-type (WT) and S100A9 knockout (KO) mice were infected with IAV (2 MOI). The medium supernatant was collected to assess levels of mouse IL-6 by ELISA. ( C ) WT and S100A9 KO BMDM were infected with IAV (2 MOI). At the indicated post-infection time-period, medium supernatant was collected to assess levels of mouse TNF-α(TNF) by ELISA. ( D ) S100A9 KO BMDMs were infected with IAV (2 MOI) in the presence of purified recombinant mouse S100A9 protein (5 µg/ml). Medium supernatant was collected from infected cells to assess levels of mouse TNF and IL-6 by ELISA. Vehicle control cells (veh) were incubated with HBSS buffer. The values represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test.

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Infection, Blocking Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Isolation, Knock-Out, Purification, Recombinant, Incubation, Standard Deviation

    ( A ) Mouse J774A.1 macrophages were incubated with purified recombinant mouse S100A9 protein (5 µg/ml) for 48 h and 72 h. The apoptotic state of these cells was examined by FACS analysis of annexin V and PI stained cells. Apoptosis rate (% apoptosis) was calculated based on number of annexin V positive/PI negative cells (denoting early apoptosis)+number of annexin V positive/PI positive cells (denoting late apoptosis)/total number of cells. ( B ) Mouse alveolar macrophage MH-S cell-line was incubated with purified S100A9 protein (5 µg/ml) for 48 h and 72 h. The apoptotic status was determined as described in (A). ( C ) Mouse J774A.1 macrophages were infected with IAV (2 MOI) in the presence of either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab). At 48 h post-infection, the apoptotic state of these cells was determined as described in (A). The values (i.e. annexin V and PI staining quantified by FACS) represents mean ± standard deviation from three independent experiments, *p<0.05 by Student's t test. Veh; cells incubated with HBSS buffer (vehicle control).

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: ( A ) Mouse J774A.1 macrophages were incubated with purified recombinant mouse S100A9 protein (5 µg/ml) for 48 h and 72 h. The apoptotic state of these cells was examined by FACS analysis of annexin V and PI stained cells. Apoptosis rate (% apoptosis) was calculated based on number of annexin V positive/PI negative cells (denoting early apoptosis)+number of annexin V positive/PI positive cells (denoting late apoptosis)/total number of cells. ( B ) Mouse alveolar macrophage MH-S cell-line was incubated with purified S100A9 protein (5 µg/ml) for 48 h and 72 h. The apoptotic status was determined as described in (A). ( C ) Mouse J774A.1 macrophages were infected with IAV (2 MOI) in the presence of either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab). At 48 h post-infection, the apoptotic state of these cells was determined as described in (A). The values (i.e. annexin V and PI staining quantified by FACS) represents mean ± standard deviation from three independent experiments, *p<0.05 by Student's t test. Veh; cells incubated with HBSS buffer (vehicle control).

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Incubation, Purification, Recombinant, Staining, Infection, Blocking Assay, Standard Deviation

    Primary bone marrow derived macrophages (BMDM) isolated from wild-type (WT) and TLR4 knockout (KO) mice were incubated with purified recombinant mouse S100A9 protein (5 ug/mL). The medium supernatant was collected to assess levels of mouse IL-6 ( A ) and mouse TNF-α(TNF) ( B ) by ELISA. ( C ) IL-6 production from S100A9 protein treated WT and MyD88 KO BMDMs. ( D ) BMDM isolated from WT, TLR4 KO and MyD88 KO mice were infected with IAV (2 MOI). At 12 h and 24 h post-infection time-period, medium supernatant was collected to assess levels of mouse IL-6 by ELISA. ( E ) TNF production from IAV infected WT and TLR4 KO BMDMs. The values represent the mean ± standard deviation from three independent experiments performed in triplicate, *p<0.05 using a Student's t test. Veh; cells incubated with HBSS buffer (vehicle control).

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: Primary bone marrow derived macrophages (BMDM) isolated from wild-type (WT) and TLR4 knockout (KO) mice were incubated with purified recombinant mouse S100A9 protein (5 ug/mL). The medium supernatant was collected to assess levels of mouse IL-6 ( A ) and mouse TNF-α(TNF) ( B ) by ELISA. ( C ) IL-6 production from S100A9 protein treated WT and MyD88 KO BMDMs. ( D ) BMDM isolated from WT, TLR4 KO and MyD88 KO mice were infected with IAV (2 MOI). At 12 h and 24 h post-infection time-period, medium supernatant was collected to assess levels of mouse IL-6 by ELISA. ( E ) TNF production from IAV infected WT and TLR4 KO BMDMs. The values represent the mean ± standard deviation from three independent experiments performed in triplicate, *p<0.05 using a Student's t test. Veh; cells incubated with HBSS buffer (vehicle control).

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Derivative Assay, Isolation, Knock-Out, Incubation, Purification, Recombinant, Enzyme-linked Immunosorbent Assay, Infection, Standard Deviation

    ( A ) Primary bone marrow derived macrophages (BMDM) isolated from wild-type (WT) and TLR4 knockout (KO) mice were incubated with purified recombinant mouse S100A9 protein (5 ug/mL) for 72 h. The apoptotic state of these cells was examined by FACS analysis of annexin V and PI stained cells. Apoptosis rate (% apoptosis) was calculated based on number of annexin V positive/PI negative cells (denoting early apoptosis)+number of annexin V positive/PI positive cells (denoting late apoptosis)/total number of cells. ( B ) WT and TLR4 KO BMDMs were infected with IAV (1 MOI). At 48 h post-infection, the apoptotic status was determined as described in (A). ( C ) IAV infected WT and TLR4 KO cells were subjected to TUNEL assay. TUNEL positive cells were analyzed by image J software. Percent TUNEL positive cells denotes ratio of number of TUNEL positive cells/total number of cells. ( D ) WT and MyD88 KO BMDMs were infected with IAV (1 MOI). At 48 h post-infection, the apoptotic status was determined. The values represents mean ± standard deviation from three independent experiments, *p<0.05 by Student's t test. Veh; cells incubated with HBSS buffer (vehicle control).

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: ( A ) Primary bone marrow derived macrophages (BMDM) isolated from wild-type (WT) and TLR4 knockout (KO) mice were incubated with purified recombinant mouse S100A9 protein (5 ug/mL) for 72 h. The apoptotic state of these cells was examined by FACS analysis of annexin V and PI stained cells. Apoptosis rate (% apoptosis) was calculated based on number of annexin V positive/PI negative cells (denoting early apoptosis)+number of annexin V positive/PI positive cells (denoting late apoptosis)/total number of cells. ( B ) WT and TLR4 KO BMDMs were infected with IAV (1 MOI). At 48 h post-infection, the apoptotic status was determined as described in (A). ( C ) IAV infected WT and TLR4 KO cells were subjected to TUNEL assay. TUNEL positive cells were analyzed by image J software. Percent TUNEL positive cells denotes ratio of number of TUNEL positive cells/total number of cells. ( D ) WT and MyD88 KO BMDMs were infected with IAV (1 MOI). At 48 h post-infection, the apoptotic status was determined. The values represents mean ± standard deviation from three independent experiments, *p<0.05 by Student's t test. Veh; cells incubated with HBSS buffer (vehicle control).

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Derivative Assay, Isolation, Knock-Out, Incubation, Purification, Recombinant, Staining, Infection, TUNEL Assay, Software, Standard Deviation

    ( A ) RNA isolated from mock infected and IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice were subjected to RT-PCR analysis to examine expression of mouse S100A9. The RT-PCR data represents three mice/group (i.e. three mock mice, three mice infected with IAV for 3 d and three mice infected with IAV for 6 d). The RT-PCR data is a representative of three independent experiments with similar results. ( B ) Lung homogenate prepared from mock infected and IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice were subjected to ELISA analysis to determine levels of mS100A9 protein in the lung. ( C ) Immuno-histochemical analysis of mouse lung tissue sections derived from mock infected and IAV infected mice were stained with mouse S100A9 antibody. Magnification, 200×. One representative example of a total of 3 mice analyzed per group in two independent experiments. ( D ) Broncho-alveolar lavage fluid (BALF) isolated from mock infected and IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice were subjected to ELISA analysis to determine levels of S100A9 protein in BALF. The values shown in (B) and (D) represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test.

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: ( A ) RNA isolated from mock infected and IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice were subjected to RT-PCR analysis to examine expression of mouse S100A9. The RT-PCR data represents three mice/group (i.e. three mock mice, three mice infected with IAV for 3 d and three mice infected with IAV for 6 d). The RT-PCR data is a representative of three independent experiments with similar results. ( B ) Lung homogenate prepared from mock infected and IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice were subjected to ELISA analysis to determine levels of mS100A9 protein in the lung. ( C ) Immuno-histochemical analysis of mouse lung tissue sections derived from mock infected and IAV infected mice were stained with mouse S100A9 antibody. Magnification, 200×. One representative example of a total of 3 mice analyzed per group in two independent experiments. ( D ) Broncho-alveolar lavage fluid (BALF) isolated from mock infected and IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice were subjected to ELISA analysis to determine levels of S100A9 protein in BALF. The values shown in (B) and (D) represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test.

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Isolation, Infection, Reverse Transcription Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Staining, Standard Deviation

    ( A ) Survival of IAV infected (1×10 5 pfu/mouse via intra-tracheal route) mice administered with either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab) (24 h prior to IAV inoculation, 2 mg of antibody/mouse administered via i.p route). The data represents values from two independent experiments performed with 5 mice/group for each experiment (total 10 mice/group from two experiments); *p = 0.03. ( B ) Hematoxylin and eosin (H&E) staining of lung sections from mock infected or IAV infected mice (3×10 4 pfu/mouse via intra-tracheal route) administered with either control IgG (IgG) or S100A9 Ab (24 h prior to IAV inoculation, 2 mg of antibody/mouse was administered via i.p route). Magnification, ×10. ( C ) Mice were administered with purified recombinant mouse S100A9 protein (15 µg/mouse) via intra-tracheal route. At 8 h post-administration, levels of mouse TNF-α in the lung was assessed by performing ELISA analysis with lung homogenate. ( D ) Lung homogenate prepared from mock infected and IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice administered with either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab) (24 h prior to IAV inoculation, 2 mg of antibody/mouse administered via i.p route) were subjected to ELISA analysis to determine levels of mouse TNF-α in the lung. ( E ) For ex-vivo experiment, broncho-alveolar lavage fluid (BALF) was collected (at 3 d post-infection) from IAV infected mice (2×10 4 pfu/mouse via intra-tracheal route) administered with either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab) (24 h prior to IAV inoculation, 2 mg of antibody/mouse administered via i.p route). The BALF cells were isolated and plated in 48-well plate. After 2 h and 4 h, the medium supernatant was analyzed for mouse TNF-α (TNF) and mouse IL-6 by ELISA. Values shown in (C), (D) and (E) represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test. Veh; HBSS buffer diluted in PBS (vehicle control).

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: ( A ) Survival of IAV infected (1×10 5 pfu/mouse via intra-tracheal route) mice administered with either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab) (24 h prior to IAV inoculation, 2 mg of antibody/mouse administered via i.p route). The data represents values from two independent experiments performed with 5 mice/group for each experiment (total 10 mice/group from two experiments); *p = 0.03. ( B ) Hematoxylin and eosin (H&E) staining of lung sections from mock infected or IAV infected mice (3×10 4 pfu/mouse via intra-tracheal route) administered with either control IgG (IgG) or S100A9 Ab (24 h prior to IAV inoculation, 2 mg of antibody/mouse was administered via i.p route). Magnification, ×10. ( C ) Mice were administered with purified recombinant mouse S100A9 protein (15 µg/mouse) via intra-tracheal route. At 8 h post-administration, levels of mouse TNF-α in the lung was assessed by performing ELISA analysis with lung homogenate. ( D ) Lung homogenate prepared from mock infected and IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice administered with either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab) (24 h prior to IAV inoculation, 2 mg of antibody/mouse administered via i.p route) were subjected to ELISA analysis to determine levels of mouse TNF-α in the lung. ( E ) For ex-vivo experiment, broncho-alveolar lavage fluid (BALF) was collected (at 3 d post-infection) from IAV infected mice (2×10 4 pfu/mouse via intra-tracheal route) administered with either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab) (24 h prior to IAV inoculation, 2 mg of antibody/mouse administered via i.p route). The BALF cells were isolated and plated in 48-well plate. After 2 h and 4 h, the medium supernatant was analyzed for mouse TNF-α (TNF) and mouse IL-6 by ELISA. Values shown in (C), (D) and (E) represent the mean ± standard deviation from three independent experiments performed in triplicate. *p<0.05 using a Student's t test. Veh; HBSS buffer diluted in PBS (vehicle control).

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Infection, Blocking Assay, Staining, Purification, Recombinant, Enzyme-linked Immunosorbent Assay, Ex Vivo, Isolation, Standard Deviation

    ( A ) Lung sections were prepared (at 3 d post-infection) from IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice administered with either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab) (24 h prior to IAV inoculation, 2 mg of antibody/mouse administered via i.p route). For each experimental group lung sections were prepared from three control IgG treated mice (+IAV) and three S100A9 Ab treated mice (+IAV). The lung sections were used for TUNEL staining. Image J software was used to calculate TUNEL-positive areas (representing apoptosis) in the lung sections as detailed in the methods section. The data is presented as percent apoptotic area. The percent apoptotic area was calculated from nine areas/lung section as detailed in the methods section. The values were compiled to calculate the percent apoptotic area in IAV infected IgG treated mice vs. IAV infected S100A9 Ab treated mice, * p = 0.0164 by Student's t test. ( B ) A representative TUNEL staining of lung sections from IAV infected mice administered with either IgG or S100A9 Ab. The apoptotic nuclei (representing apoptosis) are indicated with red arrows. ( C ) A schematic model depicting the role of extracellular S100A9 and DDX21/TRIF/S100A9/TLR4/MyD88 signaling network in exaggerating lung disease during IAV infection. PM, plasma membrane; NM, nuclear membrane.

    Journal: PLoS Pathogens

    Article Title: DAMP Molecule S100A9 Acts as a Molecular Pattern to Enhance Inflammation during Influenza A Virus Infection: Role of DDX21-TRIF-TLR4-MyD88 Pathway

    doi: 10.1371/journal.ppat.1003848

    Figure Lengend Snippet: ( A ) Lung sections were prepared (at 3 d post-infection) from IAV infected (2×10 4 pfu/mouse via intra-tracheal route) mice administered with either control IgG (IgG) or anti-S100A9 blocking (neutralizing) antibody (S100A9 Ab) (24 h prior to IAV inoculation, 2 mg of antibody/mouse administered via i.p route). For each experimental group lung sections were prepared from three control IgG treated mice (+IAV) and three S100A9 Ab treated mice (+IAV). The lung sections were used for TUNEL staining. Image J software was used to calculate TUNEL-positive areas (representing apoptosis) in the lung sections as detailed in the methods section. The data is presented as percent apoptotic area. The percent apoptotic area was calculated from nine areas/lung section as detailed in the methods section. The values were compiled to calculate the percent apoptotic area in IAV infected IgG treated mice vs. IAV infected S100A9 Ab treated mice, * p = 0.0164 by Student's t test. ( B ) A representative TUNEL staining of lung sections from IAV infected mice administered with either IgG or S100A9 Ab. The apoptotic nuclei (representing apoptosis) are indicated with red arrows. ( C ) A schematic model depicting the role of extracellular S100A9 and DDX21/TRIF/S100A9/TLR4/MyD88 signaling network in exaggerating lung disease during IAV infection. PM, plasma membrane; NM, nuclear membrane.

    Article Snippet: The plates were washed three times with PBST and incubated with either goat anti-mouse IgG (300 ng/well) (R&D) (for mouse S100A9) or mouse anti-human IgG (50 ng/well) (R&D) (for human S100A9) in PBST+0.1% BSA for 2 h at room temperature.

    Techniques: Infection, Blocking Assay, TUNEL Assay, Staining, Software