immune complex stimulated  (Thermo Fisher)


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    Structured Review

    Thermo Fisher immune complex stimulated
    Immune Complex Stimulated, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immune complex stimulated/product/Thermo Fisher
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immune complex stimulated - by Bioz Stars, 2020-07
    80/100 stars

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    Flow Cytometry:

    Article Title: Characterization of tumor necrosis factor-deficient mice
    Article Snippet: .. Respiratory burst activity was measured by flow cytometric analysis of phorbol 12-myristate 13-acetate (Sigma) or immune complex-stimulated (Molecular Probes) whole blood ( ). .. Phagocytic activity of thioglycollate-elicited macrophages for sheep red blood cells (SRBC) (Cedarlane Laboratories), or opsonized SRBC using subagglutinizing doses of anti-sheep IgG (Sigma), and the induction of nitrite were determined ( ).

    Activity Assay:

    Article Title: Characterization of tumor necrosis factor-deficient mice
    Article Snippet: .. Respiratory burst activity was measured by flow cytometric analysis of phorbol 12-myristate 13-acetate (Sigma) or immune complex-stimulated (Molecular Probes) whole blood ( ). .. Phagocytic activity of thioglycollate-elicited macrophages for sheep red blood cells (SRBC) (Cedarlane Laboratories), or opsonized SRBC using subagglutinizing doses of anti-sheep IgG (Sigma), and the induction of nitrite were determined ( ).

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    Thermo Fisher gene exp cd74 mm00658576 m1
    Preferential differentiation of IL-21 producing T cells by co-culture with Blimp-1 deficient DCs in vitro DCs (spleens of female wild-type (open symbol) or CKO (closed symbol) mice) and <t>OT-II</t> CD4 + T cells were co-cultured for 3 days at various ratios of DCs to T cells with OVA protein (10 µg/ml). (a) To assess proliferation, <t>OT-II</t> T cells were labeled with CFSE (10 µM) before co-culture with DCs. Proliferation was measured by calculation of division index (total number of divisions/number of starting cells). A representative flow image is presented on the left (red line represents wild-type DCs and filled blue represents Blimp-1 deficient DCs). The box-and-whisker plot, horizontal bars indicate the median, boxes indicate 25 th to 75 th percentile, and whiskers indicate 10 th and 90 th percentile (n=4). (b) To assess cytokine expression, T cells were harvested after co-culture with DCs (either unfixed DCs or fixed DCs as designated in figure) and total RNA was purified. Cytokine mRNA was measured by qPCR and the relative expression was calculated by normalization to Polr2a . In the box-and-whisker plots, horizontal bars indicate the median, boxes indicate 25 th to 75 th percentile, and whiskers indicate 10 th and 90 th percentile (unfixed DCs (n=12) and fixed DCs (n=10)). (c) Protein level of cytokines was measured by intracellular staining of T cells. After co-culture, T cells were stimulated with PMA (100 ng/ml)/ionomycin (1 µg/ml) for 6 hours. BFA (20 µg/ml) was added during the last 4 hours of stimulation. Cytokine-positive cells were identified by flow cytometry and their frequency was calculated. In the box-and-whisker plot, horizontal bars indicate the median, boxes indicate 25 th to 75 th percentile, and whiskers indicate 10 th and 90 th percentile (n=8). For both (b) and (c), 1 nM CTSS inhibitor was added during the OVA protein incubation with DCs or cells were cultured in transwell plates. Flow images are from one representative set from 4 individual experiments. (d) DQ-OVA protein was incubated with DCs from wild-type (open circle) or CKO (closed circle) mice overnight. After incubation, DCs were extensively washed and OVA uptake was measured by flow cytometry (upper panel, a representative image from 3 individual experiments). The Geo MFI is presented as a dot plot graph (bottom graph). Each do represents an individual experiment and the bar represents the mean ± SEM (n=3).
    Gene Exp Cd74 Mm00658576 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher immune reaction against cd3 antigen
    Decline of splenic pDCs during systemic viral infections. Mice were infected with RNA or DNA viruses and absolute numbers of pDCs and serum IFN-α levels were determined at different time points p.i. (A) C57BL/6 mice were infected with 10 7 pfu HSV-1. Data are representative of two experiments with four mice per time point. (B) C57BL/6 mice were infected with 5 × 10 6 pfu VSV-OVA. Data are representative of two experiments with four mice per time point. (C) C57BL/6 mice were infected with 5 × 10 4 pfu MCMV. Data are combined from two experiments with five to six mice per time point. (D) Spleen sections from uninfected (Ctrl) and MCMV-infected C57BL/6 mice were stained for Siglec-H (blue) and <t>CD3</t> (brown). A magnification is shown in the inset. Bar, 100 µm. The graph shows mean pDC numbers per high power field (HPF) in Ctrl and MCMV-infected spleen sections ( n = 8). (E) pDC numbers in spleens and serum IFN-α levels were determined from C57BL/6 (5 × 10 4 pfu MCMV; 10 4 pfu Δm157) and 129/SvJ (10 4 pfu MCMV) mice infected or not for 48 h ( n = 4–5 mice per group). (F) C57BL/6 mice were infected with 3 × 10 6 pfu LCMV clone 13. Data are combined from two experiments with six to eight mice per time point. (G) C57BL/6 mice were infected with 2 × 10 6 pfu VACV. Data are representative of two experiments with three to five mice per time point. (A–C and E–G) Error bars represent means ± SEM. (A, F, and G) pDC numbers, black bars; IFN-α levels, white bars. (B and C) pDC numbers, open squares; IFN-α levels, closed circles. (D) Statistical significance is indicated by **, P
    Immune Reaction Against Cd3 Antigen, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Preferential differentiation of IL-21 producing T cells by co-culture with Blimp-1 deficient DCs in vitro DCs (spleens of female wild-type (open symbol) or CKO (closed symbol) mice) and OT-II CD4 + T cells were co-cultured for 3 days at various ratios of DCs to T cells with OVA protein (10 µg/ml). (a) To assess proliferation, OT-II T cells were labeled with CFSE (10 µM) before co-culture with DCs. Proliferation was measured by calculation of division index (total number of divisions/number of starting cells). A representative flow image is presented on the left (red line represents wild-type DCs and filled blue represents Blimp-1 deficient DCs). The box-and-whisker plot, horizontal bars indicate the median, boxes indicate 25 th to 75 th percentile, and whiskers indicate 10 th and 90 th percentile (n=4). (b) To assess cytokine expression, T cells were harvested after co-culture with DCs (either unfixed DCs or fixed DCs as designated in figure) and total RNA was purified. Cytokine mRNA was measured by qPCR and the relative expression was calculated by normalization to Polr2a . In the box-and-whisker plots, horizontal bars indicate the median, boxes indicate 25 th to 75 th percentile, and whiskers indicate 10 th and 90 th percentile (unfixed DCs (n=12) and fixed DCs (n=10)). (c) Protein level of cytokines was measured by intracellular staining of T cells. After co-culture, T cells were stimulated with PMA (100 ng/ml)/ionomycin (1 µg/ml) for 6 hours. BFA (20 µg/ml) was added during the last 4 hours of stimulation. Cytokine-positive cells were identified by flow cytometry and their frequency was calculated. In the box-and-whisker plot, horizontal bars indicate the median, boxes indicate 25 th to 75 th percentile, and whiskers indicate 10 th and 90 th percentile (n=8). For both (b) and (c), 1 nM CTSS inhibitor was added during the OVA protein incubation with DCs or cells were cultured in transwell plates. Flow images are from one representative set from 4 individual experiments. (d) DQ-OVA protein was incubated with DCs from wild-type (open circle) or CKO (closed circle) mice overnight. After incubation, DCs were extensively washed and OVA uptake was measured by flow cytometry (upper panel, a representative image from 3 individual experiments). The Geo MFI is presented as a dot plot graph (bottom graph). Each do represents an individual experiment and the bar represents the mean ± SEM (n=3).

    Journal: Nature immunology

    Article Title: Increased Cathepsin S in Prdm1−/− dendritic cells alters TFH repertoire and contributes to lupus

    doi: 10.1038/ni.3793

    Figure Lengend Snippet: Preferential differentiation of IL-21 producing T cells by co-culture with Blimp-1 deficient DCs in vitro DCs (spleens of female wild-type (open symbol) or CKO (closed symbol) mice) and OT-II CD4 + T cells were co-cultured for 3 days at various ratios of DCs to T cells with OVA protein (10 µg/ml). (a) To assess proliferation, OT-II T cells were labeled with CFSE (10 µM) before co-culture with DCs. Proliferation was measured by calculation of division index (total number of divisions/number of starting cells). A representative flow image is presented on the left (red line represents wild-type DCs and filled blue represents Blimp-1 deficient DCs). The box-and-whisker plot, horizontal bars indicate the median, boxes indicate 25 th to 75 th percentile, and whiskers indicate 10 th and 90 th percentile (n=4). (b) To assess cytokine expression, T cells were harvested after co-culture with DCs (either unfixed DCs or fixed DCs as designated in figure) and total RNA was purified. Cytokine mRNA was measured by qPCR and the relative expression was calculated by normalization to Polr2a . In the box-and-whisker plots, horizontal bars indicate the median, boxes indicate 25 th to 75 th percentile, and whiskers indicate 10 th and 90 th percentile (unfixed DCs (n=12) and fixed DCs (n=10)). (c) Protein level of cytokines was measured by intracellular staining of T cells. After co-culture, T cells were stimulated with PMA (100 ng/ml)/ionomycin (1 µg/ml) for 6 hours. BFA (20 µg/ml) was added during the last 4 hours of stimulation. Cytokine-positive cells were identified by flow cytometry and their frequency was calculated. In the box-and-whisker plot, horizontal bars indicate the median, boxes indicate 25 th to 75 th percentile, and whiskers indicate 10 th and 90 th percentile (n=8). For both (b) and (c), 1 nM CTSS inhibitor was added during the OVA protein incubation with DCs or cells were cultured in transwell plates. Flow images are from one representative set from 4 individual experiments. (d) DQ-OVA protein was incubated with DCs from wild-type (open circle) or CKO (closed circle) mice overnight. After incubation, DCs were extensively washed and OVA uptake was measured by flow cytometry (upper panel, a representative image from 3 individual experiments). The Geo MFI is presented as a dot plot graph (bottom graph). Each do represents an individual experiment and the bar represents the mean ± SEM (n=3).

    Article Snippet: Antibodies and reagents Taqman primers were purchased from Applied Biosystems: Mm00515580_m1 (Ctsc ), Mm00515586_m1 (Ctsd ), Mm00456010_m1 (Ctse ), Mm00456011_m1 (Ctsg ), Mm00484039_m1 (Ctsk ), Mm00617413_m1 (Ctso ), Mm00515599_m1 (Ctsw ), Mm01255859_m1 (CtsS ), Mm00515597_m1 (Ctsl ), Mm00613524_m1 (March1 ), Mm00439211_m1 (H2-Aa ), Mm00482914_m1 (Ciita ), Mm00783707_s1 (H2-dmb2 ), Mm00511327_m1 (Zbtb46 ), Mm00658576_m1 (CD74 ), Mm00468476_m1 (H2-oa ), Mm01325350_m1 (Lgmn ), Mm00434455_m1 (Itgam ), Mm00438347_m1 (Cst3 ), Mm00499585_m1 (CD209b ), Mm00516431_m1 (Irf4 ), Mm00477633_m1 (Bcl6 ), Mm00581355_s1 (Maf ), Mm00479410_m1 (Batf ), Mm00839502_m1 (Polr2a ), and Mm03024075_m1 (Hprt ).

    Techniques: Co-Culture Assay, In Vitro, Mouse Assay, Cell Culture, Labeling, Flow Cytometry, Whisker Assay, Expressing, Purification, Real-time Polymerase Chain Reaction, Staining, Cytometry, Incubation

    CD74 expression is upregulated in EAE

    Journal: European journal of immunology

    Article Title: Partial MHC class II constructs inhibit MIF/CD74 binding and downstream effects

    doi: 10.1002/eji.201243162

    Figure Lengend Snippet: CD74 expression is upregulated in EAE

    Article Snippet: Quantitative real time PCR was performed using the ABI7000 sequence detection system with gene-on-demand assay products (Applied Biosystems) for TNF-a (Assay ID: Mm00443260_g1), IFN-g (Assay ID: Mm01168134_m1), IL-1a (Assay ID: Mm00439620_M1), IL-1b (Assay ID: Mm00434228_m1), CD74 (Assay ID: Mm00658576_m1), CCL2 (Assay ID: Mm00441242_m1), CCR2 (Assay ID: Mm00438270_m1), IL-23 (Assay ID: Mm01160011_g1), MMP-2 (Assay ID: Mm00439498_m1), MMP-9 (Assay ID: Mm00442991_m1), CASPASE-8 (Assay ID: Mm00802247_m1), and IL-6 (Assay ID: Mm00446190_m1).

    Techniques: Expressing

    RTL treatment affects CD74 expression levels and the inflammatory milieu in the CNS

    Journal: European journal of immunology

    Article Title: Partial MHC class II constructs inhibit MIF/CD74 binding and downstream effects

    doi: 10.1002/eji.201243162

    Figure Lengend Snippet: RTL treatment affects CD74 expression levels and the inflammatory milieu in the CNS

    Article Snippet: Quantitative real time PCR was performed using the ABI7000 sequence detection system with gene-on-demand assay products (Applied Biosystems) for TNF-a (Assay ID: Mm00443260_g1), IFN-g (Assay ID: Mm01168134_m1), IL-1a (Assay ID: Mm00439620_M1), IL-1b (Assay ID: Mm00434228_m1), CD74 (Assay ID: Mm00658576_m1), CCL2 (Assay ID: Mm00441242_m1), CCR2 (Assay ID: Mm00438270_m1), IL-23 (Assay ID: Mm01160011_g1), MMP-2 (Assay ID: Mm00439498_m1), MMP-9 (Assay ID: Mm00442991_m1), CASPASE-8 (Assay ID: Mm00802247_m1), and IL-6 (Assay ID: Mm00446190_m1).

    Techniques: Expressing

    Increased expression of CD74 in MS subjects is significantly reduced by the RTL1000 construct

    Journal: European journal of immunology

    Article Title: Partial MHC class II constructs inhibit MIF/CD74 binding and downstream effects

    doi: 10.1002/eji.201243162

    Figure Lengend Snippet: Increased expression of CD74 in MS subjects is significantly reduced by the RTL1000 construct

    Article Snippet: Quantitative real time PCR was performed using the ABI7000 sequence detection system with gene-on-demand assay products (Applied Biosystems) for TNF-a (Assay ID: Mm00443260_g1), IFN-g (Assay ID: Mm01168134_m1), IL-1a (Assay ID: Mm00439620_M1), IL-1b (Assay ID: Mm00434228_m1), CD74 (Assay ID: Mm00658576_m1), CCL2 (Assay ID: Mm00441242_m1), CCR2 (Assay ID: Mm00438270_m1), IL-23 (Assay ID: Mm01160011_g1), MMP-2 (Assay ID: Mm00439498_m1), MMP-9 (Assay ID: Mm00442991_m1), CASPASE-8 (Assay ID: Mm00802247_m1), and IL-6 (Assay ID: Mm00446190_m1).

    Techniques: Expressing, Mass Spectrometry, Construct

    RTL342M blocks binding of rhMIF to CD74 immunoprecipitated from DR*1501-Tg mouse splenocytes

    Journal: European journal of immunology

    Article Title: Partial MHC class II constructs inhibit MIF/CD74 binding and downstream effects

    doi: 10.1002/eji.201243162

    Figure Lengend Snippet: RTL342M blocks binding of rhMIF to CD74 immunoprecipitated from DR*1501-Tg mouse splenocytes

    Article Snippet: Quantitative real time PCR was performed using the ABI7000 sequence detection system with gene-on-demand assay products (Applied Biosystems) for TNF-a (Assay ID: Mm00443260_g1), IFN-g (Assay ID: Mm01168134_m1), IL-1a (Assay ID: Mm00439620_M1), IL-1b (Assay ID: Mm00434228_m1), CD74 (Assay ID: Mm00658576_m1), CCL2 (Assay ID: Mm00441242_m1), CCR2 (Assay ID: Mm00438270_m1), IL-23 (Assay ID: Mm01160011_g1), MMP-2 (Assay ID: Mm00439498_m1), MMP-9 (Assay ID: Mm00442991_m1), CASPASE-8 (Assay ID: Mm00802247_m1), and IL-6 (Assay ID: Mm00446190_m1).

    Techniques: Binding Assay, Immunoprecipitation

    PIM overexpression correlates with an immune system-dependent genes signature in human breast, endometrium and ovary tumors ( A ) The figure shows a heat map comparing immunology-related genes with PIM1 and PIM2 (See Supplementary Table 3 ) in breast tumors (Halfwerk dataset, R2). Heat map shows a clear correlation of PIM1 and PIM2 with the expression of antigen presentation genes. ( B ) The figure shows a heat map comparing immunology-related genes with PIM1 and PIM2 (See Supplementary Table 4 ) in endometrial tumors (EXPO dataset, R2). Heat map shows a clear correlation of PIM1 and PIM2 with the expression of antigen presentation genes. ( C ) The figure shows a heat map comparing immunology-related genes with PIM1 and PIM2 (See Supplementary Table 5 ) in ovarian tumors (Wong dataset, R2). Heat map shows a clear correlation of PIM1 and PIM2 with the expression of antigen presentation genes. ( D ) A Venn diagram showing the number of immune system-dependent genes that correlated with PIM1 and PIM2 overexpression in Breast, endometrial and ovary tumors. ( E ) A list of system-dependent genes in Breast, endometrial and ovary tumors that correlated with both PIM1 and PIM2 overexpression (common) and that was unique to each tumor type. ( F , G ) Pro-inflammatory microenvironment PIM-dependent characterization by qPCR. The figure shows a number of immune system-dependent genes that were correlated with PIM overexpression in breast, endometrium and ovarian datasets: CD4, CD8a, CD74, Ifng, Tap1 and Tap2; and mouse Major Histocompatibility Complex markers (H2 complex). We decided to focus on the ones which has a mouse ortholog: HLA-DMA (H2-DMa), HLA-DRA (H2-A), HLA-DPB1 (CD74), and also H2-K (classical MHC class I), H2-Q (non-classical MHC class I), H2-E (classical MHC class II). They were measured by qRT-PCR of mammary gland of WT mice with non-proliferative status (MG-N) and carcinoma grades (MG-T) in both transgenic models (F), and the same way in uterus (UT-N, normal; and UT-T, carcinoma) (G). ( H ) The overexpression levels of the human transgenes PIM1 or PIM2 in the models causes an increase in the expression of NFAT and NFAT effectors (OSM and IL6), as well as the phosphorylation of mTOR. The p -value was obtained using a one-tailed student's t -test. (* p

    Journal: Oncotarget

    Article Title: Inflammation and stem markers association to PIM1/PIM2 kinase-induced tumors in breast and uterus

    doi: 10.18632/oncotarget.19438

    Figure Lengend Snippet: PIM overexpression correlates with an immune system-dependent genes signature in human breast, endometrium and ovary tumors ( A ) The figure shows a heat map comparing immunology-related genes with PIM1 and PIM2 (See Supplementary Table 3 ) in breast tumors (Halfwerk dataset, R2). Heat map shows a clear correlation of PIM1 and PIM2 with the expression of antigen presentation genes. ( B ) The figure shows a heat map comparing immunology-related genes with PIM1 and PIM2 (See Supplementary Table 4 ) in endometrial tumors (EXPO dataset, R2). Heat map shows a clear correlation of PIM1 and PIM2 with the expression of antigen presentation genes. ( C ) The figure shows a heat map comparing immunology-related genes with PIM1 and PIM2 (See Supplementary Table 5 ) in ovarian tumors (Wong dataset, R2). Heat map shows a clear correlation of PIM1 and PIM2 with the expression of antigen presentation genes. ( D ) A Venn diagram showing the number of immune system-dependent genes that correlated with PIM1 and PIM2 overexpression in Breast, endometrial and ovary tumors. ( E ) A list of system-dependent genes in Breast, endometrial and ovary tumors that correlated with both PIM1 and PIM2 overexpression (common) and that was unique to each tumor type. ( F , G ) Pro-inflammatory microenvironment PIM-dependent characterization by qPCR. The figure shows a number of immune system-dependent genes that were correlated with PIM overexpression in breast, endometrium and ovarian datasets: CD4, CD8a, CD74, Ifng, Tap1 and Tap2; and mouse Major Histocompatibility Complex markers (H2 complex). We decided to focus on the ones which has a mouse ortholog: HLA-DMA (H2-DMa), HLA-DRA (H2-A), HLA-DPB1 (CD74), and also H2-K (classical MHC class I), H2-Q (non-classical MHC class I), H2-E (classical MHC class II). They were measured by qRT-PCR of mammary gland of WT mice with non-proliferative status (MG-N) and carcinoma grades (MG-T) in both transgenic models (F), and the same way in uterus (UT-N, normal; and UT-T, carcinoma) (G). ( H ) The overexpression levels of the human transgenes PIM1 or PIM2 in the models causes an increase in the expression of NFAT and NFAT effectors (OSM and IL6), as well as the phosphorylation of mTOR. The p -value was obtained using a one-tailed student's t -test. (* p

    Article Snippet: We used the following TaqMan Gene Expression Assays probes (ThermoFisher) after RT-PCR: PIM1 (Hs01065498_m1), PIM2 (Hs00179139_m1), Pim1 (Mm00435712_m1), Pim2 (Mm00454579_m1), Meox1 (Mm00440285_m1), Sox15 (Mm00839542_g1), Pou5f1 (Mm03053917_g1), Nanog (Mm02019550_s1), Sox2 (Mm03053810_s1), Klf4 (Mm00516104_m1), Klf10 (Mm00449812_m1), Serpine1 (Mm00435858_m1), Thbd (Mm00437014_s1), CD4 (Mm00442754_m1), CD8a (Mm01182107_g1), CD74 (Mm00658576_m1), Ifng (Mm01168134_m1), Tap1 (Mm00443188_m1), Tap2 (Mm01277033_m1).

    Techniques: Over Expression, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Mouse Assay, Transgenic Assay, One-tailed Test

    Decline of splenic pDCs during systemic viral infections. Mice were infected with RNA or DNA viruses and absolute numbers of pDCs and serum IFN-α levels were determined at different time points p.i. (A) C57BL/6 mice were infected with 10 7 pfu HSV-1. Data are representative of two experiments with four mice per time point. (B) C57BL/6 mice were infected with 5 × 10 6 pfu VSV-OVA. Data are representative of two experiments with four mice per time point. (C) C57BL/6 mice were infected with 5 × 10 4 pfu MCMV. Data are combined from two experiments with five to six mice per time point. (D) Spleen sections from uninfected (Ctrl) and MCMV-infected C57BL/6 mice were stained for Siglec-H (blue) and CD3 (brown). A magnification is shown in the inset. Bar, 100 µm. The graph shows mean pDC numbers per high power field (HPF) in Ctrl and MCMV-infected spleen sections ( n = 8). (E) pDC numbers in spleens and serum IFN-α levels were determined from C57BL/6 (5 × 10 4 pfu MCMV; 10 4 pfu Δm157) and 129/SvJ (10 4 pfu MCMV) mice infected or not for 48 h ( n = 4–5 mice per group). (F) C57BL/6 mice were infected with 3 × 10 6 pfu LCMV clone 13. Data are combined from two experiments with six to eight mice per time point. (G) C57BL/6 mice were infected with 2 × 10 6 pfu VACV. Data are representative of two experiments with three to five mice per time point. (A–C and E–G) Error bars represent means ± SEM. (A, F, and G) pDC numbers, black bars; IFN-α levels, white bars. (B and C) pDC numbers, open squares; IFN-α levels, closed circles. (D) Statistical significance is indicated by **, P

    Journal: The Journal of Experimental Medicine

    Article Title: Type I interferon negatively controls plasmacytoid dendritic cell numbers in vivo

    doi: 10.1084/jem.20110654

    Figure Lengend Snippet: Decline of splenic pDCs during systemic viral infections. Mice were infected with RNA or DNA viruses and absolute numbers of pDCs and serum IFN-α levels were determined at different time points p.i. (A) C57BL/6 mice were infected with 10 7 pfu HSV-1. Data are representative of two experiments with four mice per time point. (B) C57BL/6 mice were infected with 5 × 10 6 pfu VSV-OVA. Data are representative of two experiments with four mice per time point. (C) C57BL/6 mice were infected with 5 × 10 4 pfu MCMV. Data are combined from two experiments with five to six mice per time point. (D) Spleen sections from uninfected (Ctrl) and MCMV-infected C57BL/6 mice were stained for Siglec-H (blue) and CD3 (brown). A magnification is shown in the inset. Bar, 100 µm. The graph shows mean pDC numbers per high power field (HPF) in Ctrl and MCMV-infected spleen sections ( n = 8). (E) pDC numbers in spleens and serum IFN-α levels were determined from C57BL/6 (5 × 10 4 pfu MCMV; 10 4 pfu Δm157) and 129/SvJ (10 4 pfu MCMV) mice infected or not for 48 h ( n = 4–5 mice per group). (F) C57BL/6 mice were infected with 3 × 10 6 pfu LCMV clone 13. Data are combined from two experiments with six to eight mice per time point. (G) C57BL/6 mice were infected with 2 × 10 6 pfu VACV. Data are representative of two experiments with three to five mice per time point. (A–C and E–G) Error bars represent means ± SEM. (A, F, and G) pDC numbers, black bars; IFN-α levels, white bars. (B and C) pDC numbers, open squares; IFN-α levels, closed circles. (D) Statistical significance is indicated by **, P

    Article Snippet: The second immune reaction against CD3 antigen (rabbit monoclonal, clone SP7, dilution 1:100; Thermo Fisher Scientific) was visualized using Envision Rabbit-HRP (Dako) followed by DAB.

    Techniques: Mouse Assay, Infection, Staining