brefeldin a Search Results


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  • 99
    Thermo Fisher brefeldin a
    Brefeldin A, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8963 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore brefeldin a
    Brefeldin A, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 33791 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend brefeldin a
    Brefeldin A, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 3716 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson brefeldin a
    Subdominant HSV-1 epitopes expand to accommodate the loss of the immunodominant gB 498-505 epitope during acute infiltration into the TG. B6 mice received corneal infections with (A, B, D) HSV-1 WT, S1L, or (C) L8A. TG were excised at 8 dpi, dispersed into single cell suspensions, stimulated for 6 hrs in the presence of <t>Brefeldin</t> A with B6WT3 cells pulsed with peptides corresponding to known HSV-specific CD8 + T cell epitopes, stained for surface CD45, CD8, and intracellular IFNγ. (A) The graph shows the percent of the total CD8 + T cell population staining for intracellular IFNγ by flow cytometry. The bars represent the mean ± SEM frequency of CD8 + T cells producing IFNγ in response to each epitope. (B) Total fraction of gB 498-505 or non-gB-CD8s responding to peptide stimulations as seen in (A). N = 3–8 TG equivalents per peptide. (C) A fraction of the peptide library was analyzed as in (A), but in mice infected with HSV-1 L8A. (D) Spleens were also excised at 8 and 20 dpi and examined for RR1 982-989 tetramer positive cells. Shown is example of tetramer staining, and a graph depicting the total fraction of splenic CD8+ T cells that stained positive for tetramer. A t-test was performed for each matched pair of responding CD8 + T cells, and * denotes p
    Brefeldin A, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 15653 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher permeabilization
    Subdominant HSV-1 epitopes expand to accommodate the loss of the immunodominant gB 498-505 epitope during acute infiltration into the TG. B6 mice received corneal infections with (A, B, D) HSV-1 WT, S1L, or (C) L8A. TG were excised at 8 dpi, dispersed into single cell suspensions, stimulated for 6 hrs in the presence of <t>Brefeldin</t> A with B6WT3 cells pulsed with peptides corresponding to known HSV-specific CD8 + T cell epitopes, stained for surface CD45, CD8, and intracellular IFNγ. (A) The graph shows the percent of the total CD8 + T cell population staining for intracellular IFNγ by flow cytometry. The bars represent the mean ± SEM frequency of CD8 + T cells producing IFNγ in response to each epitope. (B) Total fraction of gB 498-505 or non-gB-CD8s responding to peptide stimulations as seen in (A). N = 3–8 TG equivalents per peptide. (C) A fraction of the peptide library was analyzed as in (A), but in mice infected with HSV-1 L8A. (D) Spleens were also excised at 8 and 20 dpi and examined for RR1 982-989 tetramer positive cells. Shown is example of tetramer staining, and a graph depicting the total fraction of splenic CD8+ T cells that stained positive for tetramer. A t-test was performed for each matched pair of responding CD8 + T cells, and * denotes p
    Permeabilization, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11625 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Pharmingen brefeldin a
    Mφ proinflammatory cytokine secretion following challenge with  N. meningitidis . (a) WT and SR-A −/−  BMMφ were incubated with ethanol-fixed MC58 bacteria at 37°C. At different times, the culture supernatant was harvested and analyzed by cytokine ELISA for production of TNF-α, IL-6, IL-10, and IL-12. The results are representative of at least two similar experiments, and the average of triplicate conditions is plotted. (b) BMMφ were incubated with ethanol-fixed RdGnX-MC58 (40 bacteria per cell) in the presence of brefeldin A. After incubation, the Mφ were fixed with 4% paraformaldehyde, stained with a phycoerythrin (PE)-labeled anti-TNF-α antibody, and analyzed by flow cytometry. The results are representative of at least two similar experiments. Note that preincubation of the anti-TNF-α antibody with recombinant mouse TNF-α or of the cells with an unlabeled anti-TNF-α antibody blocked the binding of the PE-labeled anti-TNF-α (not shown), confirming specificity. The error bars indicate standard deviations.
    Brefeldin A, supplied by Pharmingen, used in various techniques. Bioz Stars score: 92/100, based on 462 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc brefeldin a
    Similar APP expression pattern by brefeldin A, a protein ER-to-Golgi traffic blocker. (A) Chemical structures of isoginkgetin and brefeldin A. (B) Western blots of time-coursed APP expression in HEK293T cells treated with 30 μM IGK, 5 μM brefeldin A, and DMSO.
    Brefeldin A, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 217 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend cell activation cocktail
    Similar APP expression pattern by brefeldin A, a protein ER-to-Golgi traffic blocker. (A) Chemical structures of isoginkgetin and brefeldin A. (B) Western blots of time-coursed APP expression in HEK293T cells treated with 30 μM IGK, 5 μM brefeldin A, and DMSO.
    Cell Activation Cocktail, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 370 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher brefeldin a solution
    Similar APP expression pattern by brefeldin A, a protein ER-to-Golgi traffic blocker. (A) Chemical structures of isoginkgetin and brefeldin A. (B) Western blots of time-coursed APP expression in HEK293T cells treated with 30 μM IGK, 5 μM brefeldin A, and DMSO.
    Brefeldin A Solution, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 365 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Enzo Biochem brefeldin a
    Similar APP expression pattern by brefeldin A, a protein ER-to-Golgi traffic blocker. (A) Chemical structures of isoginkgetin and brefeldin A. (B) Western blots of time-coursed APP expression in HEK293T cells treated with 30 μM IGK, 5 μM brefeldin A, and DMSO.
    Brefeldin A, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 97/100, based on 282 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cayman Chemical brefeldin a
    Similar APP expression pattern by brefeldin A, a protein ER-to-Golgi traffic blocker. (A) Chemical structures of isoginkgetin and brefeldin A. (B) Western blots of time-coursed APP expression in HEK293T cells treated with 30 μM IGK, 5 μM brefeldin A, and DMSO.
    Brefeldin A, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 99/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Tocris brefeldin a
    Similar APP expression pattern by brefeldin A, a protein ER-to-Golgi traffic blocker. (A) Chemical structures of isoginkgetin and brefeldin A. (B) Western blots of time-coursed APP expression in HEK293T cells treated with 30 μM IGK, 5 μM brefeldin A, and DMSO.
    Brefeldin A, supplied by Tocris, used in various techniques. Bioz Stars score: 95/100, based on 84 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Applichem brefeldin a
    The absence of Tregs or IL-10 does not compromise persistence of strain 101 in the oral epithelium.  (A–C)  DEREG mice and control littermates were sublingually infected with  C. albicans  strain 101 and treated with diphtheria toxin on day 11 and 13 after the antifungal response was fully established. One day after the last treatment, the mice were sacrificed for analysis.  (A)  Treg depletion efficiency was analyzed in the cervical lymph nodes by flow cytometry. Data are the % of Foxp3 +  cells within the population of CD4 +  viable cells.  (B)  Lymph node cells were re-stimulated with MutuDC1940 cells that were pulsed with heat-killed  C. albicans  or left unpulsed for 5 h in the presence of Brefeldin A. IL-17A (left) and IFN-γ (right) production by CD3 + CD4 +  cells was analyzed by intracellular cytokine staining and flow cytometry.  (C)  The fungal burden was determined by plating tongue homogenates on YPD agar. Each bar represents the mean + SD of 3 to 4 mice per group. Data are from one out of two independent experiments.  (D,E)  IL-10-deficient mice and WT controls were sublingually infected with  C. albicans  strain 101 and analyzed on day 9 post-infection.  (D)  Lymph node cells were re-stimulated and analyzed for IL-17 (left) and IFN-γ (right) production as in B.  (E)  Tongue fungal burdens were analyzed as in C. Each bar represents the mean + SD of 8–9 mice per group pooled from two independent experiments. Statistics were calculated using unpaired t-Test. *** p
    Brefeldin A, supplied by Applichem, used in various techniques. Bioz Stars score: 92/100, based on 147 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    N/A
    Brefeldin A has been shown to induce apoptosis in human tumor cells through caspase activation It has also been shown to block translocation of proteins from the endoplasmic reticulum to
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    N/A
    Brefeldin A BFA is a natural fungal metabolite which has been used extensively to study intracellular transport by vesicles or endosomes Early studies demonstrated that BFA reversibly interferes with protein
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    N/A
    Brefeldin A is a fungal metabolite that disrupts the structure and function of the Golgi apparatus Brefeldin A induces the Golgi proteins to redistribute into the ER and blocks the
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    Image Search Results


    Subdominant HSV-1 epitopes expand to accommodate the loss of the immunodominant gB 498-505 epitope during acute infiltration into the TG. B6 mice received corneal infections with (A, B, D) HSV-1 WT, S1L, or (C) L8A. TG were excised at 8 dpi, dispersed into single cell suspensions, stimulated for 6 hrs in the presence of Brefeldin A with B6WT3 cells pulsed with peptides corresponding to known HSV-specific CD8 + T cell epitopes, stained for surface CD45, CD8, and intracellular IFNγ. (A) The graph shows the percent of the total CD8 + T cell population staining for intracellular IFNγ by flow cytometry. The bars represent the mean ± SEM frequency of CD8 + T cells producing IFNγ in response to each epitope. (B) Total fraction of gB 498-505 or non-gB-CD8s responding to peptide stimulations as seen in (A). N = 3–8 TG equivalents per peptide. (C) A fraction of the peptide library was analyzed as in (A), but in mice infected with HSV-1 L8A. (D) Spleens were also excised at 8 and 20 dpi and examined for RR1 982-989 tetramer positive cells. Shown is example of tetramer staining, and a graph depicting the total fraction of splenic CD8+ T cells that stained positive for tetramer. A t-test was performed for each matched pair of responding CD8 + T cells, and * denotes p

    Journal: PLoS Pathogens

    Article Title: Influence of an immunodominant herpes simplex virus type 1 CD8+ T cell epitope on the target hierarchy and function of subdominant CD8+ T cells

    doi: 10.1371/journal.ppat.1006732

    Figure Lengend Snippet: Subdominant HSV-1 epitopes expand to accommodate the loss of the immunodominant gB 498-505 epitope during acute infiltration into the TG. B6 mice received corneal infections with (A, B, D) HSV-1 WT, S1L, or (C) L8A. TG were excised at 8 dpi, dispersed into single cell suspensions, stimulated for 6 hrs in the presence of Brefeldin A with B6WT3 cells pulsed with peptides corresponding to known HSV-specific CD8 + T cell epitopes, stained for surface CD45, CD8, and intracellular IFNγ. (A) The graph shows the percent of the total CD8 + T cell population staining for intracellular IFNγ by flow cytometry. The bars represent the mean ± SEM frequency of CD8 + T cells producing IFNγ in response to each epitope. (B) Total fraction of gB 498-505 or non-gB-CD8s responding to peptide stimulations as seen in (A). N = 3–8 TG equivalents per peptide. (C) A fraction of the peptide library was analyzed as in (A), but in mice infected with HSV-1 L8A. (D) Spleens were also excised at 8 and 20 dpi and examined for RR1 982-989 tetramer positive cells. Shown is example of tetramer staining, and a graph depicting the total fraction of splenic CD8+ T cells that stained positive for tetramer. A t-test was performed for each matched pair of responding CD8 + T cells, and * denotes p

    Article Snippet: The dispersed TG or spleen cells were added to peptide-pulsed or infected fibroblasts in the presence of Brefeldin A and anti-CD107a (BD clone 1D4B) for 6 hr at 37°C/5% CO2 .

    Techniques: Mouse Assay, Staining, Flow Cytometry, Cytometry, Infection

    Stimulation of acute TG-resident CD8 +  T cell populations with WT, SIL, or L8A gB peptides. B6 mice received corneal infections with HSV-1 expressing WT, S1L, or L8A gB. TG were obtained at 8 dpi, dispersed into single cell suspensions, and the endogenous CD8 +  T cells were stimulated for 6 hours with B6WT3 fibroblasts pulsed individually with WT, S1L, or L8A gB 498-505  peptides, in the presence of brefeldin A. Cells were surface stained for CD45 and CD8, followed by an intracellular stain for IFNγ. The data are represented as the mean percentage of CD8 +  T cells that produced IFNγ +/- SEM (n = 5 mice per group). * represents significance of p

    Journal: PLoS Pathogens

    Article Title: Influence of an immunodominant herpes simplex virus type 1 CD8+ T cell epitope on the target hierarchy and function of subdominant CD8+ T cells

    doi: 10.1371/journal.ppat.1006732

    Figure Lengend Snippet: Stimulation of acute TG-resident CD8 + T cell populations with WT, SIL, or L8A gB peptides. B6 mice received corneal infections with HSV-1 expressing WT, S1L, or L8A gB. TG were obtained at 8 dpi, dispersed into single cell suspensions, and the endogenous CD8 + T cells were stimulated for 6 hours with B6WT3 fibroblasts pulsed individually with WT, S1L, or L8A gB 498-505 peptides, in the presence of brefeldin A. Cells were surface stained for CD45 and CD8, followed by an intracellular stain for IFNγ. The data are represented as the mean percentage of CD8 + T cells that produced IFNγ +/- SEM (n = 5 mice per group). * represents significance of p

    Article Snippet: The dispersed TG or spleen cells were added to peptide-pulsed or infected fibroblasts in the presence of Brefeldin A and anti-CD107a (BD clone 1D4B) for 6 hr at 37°C/5% CO2 .

    Techniques: Mouse Assay, Expressing, Staining, Produced

    The CD8 +  T cell population in S1L infected TG contract more rapidly and contain a higher frequency of active non-gB-CD8s. B6 mice received corneal infections with either WT or the S1L mutant at 1x10 5  PFU/cornea. TGs were harvested at 8, 12, 16, 20, or 30 dpi and:  (A)  stained for CD45, CD3, and CD8, analyzed by flow cytometry, and data recorded as the mean number of CD8 +  T cells/TG; or stimulated for 6 hrs with  (B)  HSV-1 gB-null-EGFP infected or  (C)  PRV-gB infected B6WT3 fibroblasts in the presence of Brefeldin A. The cells were then stained for surface CD45, CD3, and CD8 and for intracellular IFNγ. Data in B and C are presented as the mean ± SEM frequency of IFNγ +  CD8 +  T cells in each TG as a fraction of total CD8 +  T cells. * p

    Journal: PLoS Pathogens

    Article Title: Influence of an immunodominant herpes simplex virus type 1 CD8+ T cell epitope on the target hierarchy and function of subdominant CD8+ T cells

    doi: 10.1371/journal.ppat.1006732

    Figure Lengend Snippet: The CD8 + T cell population in S1L infected TG contract more rapidly and contain a higher frequency of active non-gB-CD8s. B6 mice received corneal infections with either WT or the S1L mutant at 1x10 5 PFU/cornea. TGs were harvested at 8, 12, 16, 20, or 30 dpi and: (A) stained for CD45, CD3, and CD8, analyzed by flow cytometry, and data recorded as the mean number of CD8 + T cells/TG; or stimulated for 6 hrs with (B) HSV-1 gB-null-EGFP infected or (C) PRV-gB infected B6WT3 fibroblasts in the presence of Brefeldin A. The cells were then stained for surface CD45, CD3, and CD8 and for intracellular IFNγ. Data in B and C are presented as the mean ± SEM frequency of IFNγ + CD8 + T cells in each TG as a fraction of total CD8 + T cells. * p

    Article Snippet: The dispersed TG or spleen cells were added to peptide-pulsed or infected fibroblasts in the presence of Brefeldin A and anti-CD107a (BD clone 1D4B) for 6 hr at 37°C/5% CO2 .

    Techniques: Infection, Mouse Assay, Mutagenesis, Staining, Flow Cytometry, Cytometry

    Mutant gB proteins and recognition by gB-CD8s. Untreated B6WT3 cells (mock) or cells transfected with plasmids to express WT gB or gB 498-505  epitope mutants were incubated for eighteen hours. The labeling of the mutations is as depicted in   Table 1 ; WT is a plasmid which went through the mutagenesis procedure without changes. Cells were harvested for expression analysis by immunoblotting using a monoclonal gB-specific antibody (lower panel) or, in parallel, transfected cells were combined with 5x10 4  gB-CD8s from an endogenously expanded clone and stimulated for 5 h in the presence of Brefeldin A. gB-CD8s were surface stained for CD45 and CD8, permeabilized, and stained for intracellular IFNγ. The graph depicts one of two representative experiments, with the mean percent of IFNγ +  cells ( n  = 2/group) and standard error of the mean (SEM) for each stimulation.

    Journal: PLoS Pathogens

    Article Title: Influence of an immunodominant herpes simplex virus type 1 CD8+ T cell epitope on the target hierarchy and function of subdominant CD8+ T cells

    doi: 10.1371/journal.ppat.1006732

    Figure Lengend Snippet: Mutant gB proteins and recognition by gB-CD8s. Untreated B6WT3 cells (mock) or cells transfected with plasmids to express WT gB or gB 498-505 epitope mutants were incubated for eighteen hours. The labeling of the mutations is as depicted in Table 1 ; WT is a plasmid which went through the mutagenesis procedure without changes. Cells were harvested for expression analysis by immunoblotting using a monoclonal gB-specific antibody (lower panel) or, in parallel, transfected cells were combined with 5x10 4 gB-CD8s from an endogenously expanded clone and stimulated for 5 h in the presence of Brefeldin A. gB-CD8s were surface stained for CD45 and CD8, permeabilized, and stained for intracellular IFNγ. The graph depicts one of two representative experiments, with the mean percent of IFNγ + cells ( n = 2/group) and standard error of the mean (SEM) for each stimulation.

    Article Snippet: The dispersed TG or spleen cells were added to peptide-pulsed or infected fibroblasts in the presence of Brefeldin A and anti-CD107a (BD clone 1D4B) for 6 hr at 37°C/5% CO2 .

    Techniques: Mutagenesis, Transfection, Incubation, Labeling, Plasmid Preparation, Expressing, Staining

    Mφ proinflammatory cytokine secretion following challenge with  N. meningitidis . (a) WT and SR-A −/−  BMMφ were incubated with ethanol-fixed MC58 bacteria at 37°C. At different times, the culture supernatant was harvested and analyzed by cytokine ELISA for production of TNF-α, IL-6, IL-10, and IL-12. The results are representative of at least two similar experiments, and the average of triplicate conditions is plotted. (b) BMMφ were incubated with ethanol-fixed RdGnX-MC58 (40 bacteria per cell) in the presence of brefeldin A. After incubation, the Mφ were fixed with 4% paraformaldehyde, stained with a phycoerythrin (PE)-labeled anti-TNF-α antibody, and analyzed by flow cytometry. The results are representative of at least two similar experiments. Note that preincubation of the anti-TNF-α antibody with recombinant mouse TNF-α or of the cells with an unlabeled anti-TNF-α antibody blocked the binding of the PE-labeled anti-TNF-α (not shown), confirming specificity. The error bars indicate standard deviations.

    Journal: Infection and Immunity

    Article Title: The Class A Macrophage Scavenger Receptor Is a Major Pattern Recognition Receptor for Neisseria meningitidis Which Is Independent of Lipopolysaccharide and Not Required for Secretory Responses

    doi: 10.1128/IAI.70.10.5346-5354.2002

    Figure Lengend Snippet: Mφ proinflammatory cytokine secretion following challenge with N. meningitidis . (a) WT and SR-A −/− BMMφ were incubated with ethanol-fixed MC58 bacteria at 37°C. At different times, the culture supernatant was harvested and analyzed by cytokine ELISA for production of TNF-α, IL-6, IL-10, and IL-12. The results are representative of at least two similar experiments, and the average of triplicate conditions is plotted. (b) BMMφ were incubated with ethanol-fixed RdGnX-MC58 (40 bacteria per cell) in the presence of brefeldin A. After incubation, the Mφ were fixed with 4% paraformaldehyde, stained with a phycoerythrin (PE)-labeled anti-TNF-α antibody, and analyzed by flow cytometry. The results are representative of at least two similar experiments. Note that preincubation of the anti-TNF-α antibody with recombinant mouse TNF-α or of the cells with an unlabeled anti-TNF-α antibody blocked the binding of the PE-labeled anti-TNF-α (not shown), confirming specificity. The error bars indicate standard deviations.

    Article Snippet: Cytokine detection using flow cytometry kits, brefeldin A, and enzyme-linked immunosorbent assay (ELISA) kits were obtained from PharMingen (San Diego, Calif.), and brain heart infusion broth was from Merck (Poole, United Kingdom).

    Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Staining, Labeling, Flow Cytometry, Cytometry, Recombinant, Binding Assay

    Similar APP expression pattern by brefeldin A, a protein ER-to-Golgi traffic blocker. (A) Chemical structures of isoginkgetin and brefeldin A. (B) Western blots of time-coursed APP expression in HEK293T cells treated with 30 μM IGK, 5 μM brefeldin A, and DMSO.

    Journal: ACS Omega

    Article Title: Effects of RNA Splicing Inhibitors on Amyloid Precursor Protein Expression

    doi: 10.1021/acsomega.7b02073

    Figure Lengend Snippet: Similar APP expression pattern by brefeldin A, a protein ER-to-Golgi traffic blocker. (A) Chemical structures of isoginkgetin and brefeldin A. (B) Western blots of time-coursed APP expression in HEK293T cells treated with 30 μM IGK, 5 μM brefeldin A, and DMSO.

    Article Snippet: Chemicals used in other experiments were CHX (239764, Calbiochem), bafilomycin A1 (B1793, Sigma-Aldrich), MG-132 (1748, Boston Biochem), and brefeldin A (9972S, Cell Signaling Technology).

    Techniques: Expressing, Western Blot

    The absence of Tregs or IL-10 does not compromise persistence of strain 101 in the oral epithelium.  (A–C)  DEREG mice and control littermates were sublingually infected with  C. albicans  strain 101 and treated with diphtheria toxin on day 11 and 13 after the antifungal response was fully established. One day after the last treatment, the mice were sacrificed for analysis.  (A)  Treg depletion efficiency was analyzed in the cervical lymph nodes by flow cytometry. Data are the % of Foxp3 +  cells within the population of CD4 +  viable cells.  (B)  Lymph node cells were re-stimulated with MutuDC1940 cells that were pulsed with heat-killed  C. albicans  or left unpulsed for 5 h in the presence of Brefeldin A. IL-17A (left) and IFN-γ (right) production by CD3 + CD4 +  cells was analyzed by intracellular cytokine staining and flow cytometry.  (C)  The fungal burden was determined by plating tongue homogenates on YPD agar. Each bar represents the mean + SD of 3 to 4 mice per group. Data are from one out of two independent experiments.  (D,E)  IL-10-deficient mice and WT controls were sublingually infected with  C. albicans  strain 101 and analyzed on day 9 post-infection.  (D)  Lymph node cells were re-stimulated and analyzed for IL-17 (left) and IFN-γ (right) production as in B.  (E)  Tongue fungal burdens were analyzed as in C. Each bar represents the mean + SD of 8–9 mice per group pooled from two independent experiments. Statistics were calculated using unpaired t-Test. *** p

    Journal: Frontiers in Immunology

    Article Title: Persistence of Candida albicans in the Oral Mucosa Induces a Curbed Inflammatory Host Response That Is Independent of Immunosuppression

    doi: 10.3389/fimmu.2019.00330

    Figure Lengend Snippet: The absence of Tregs or IL-10 does not compromise persistence of strain 101 in the oral epithelium. (A–C) DEREG mice and control littermates were sublingually infected with C. albicans strain 101 and treated with diphtheria toxin on day 11 and 13 after the antifungal response was fully established. One day after the last treatment, the mice were sacrificed for analysis. (A) Treg depletion efficiency was analyzed in the cervical lymph nodes by flow cytometry. Data are the % of Foxp3 + cells within the population of CD4 + viable cells. (B) Lymph node cells were re-stimulated with MutuDC1940 cells that were pulsed with heat-killed C. albicans or left unpulsed for 5 h in the presence of Brefeldin A. IL-17A (left) and IFN-γ (right) production by CD3 + CD4 + cells was analyzed by intracellular cytokine staining and flow cytometry. (C) The fungal burden was determined by plating tongue homogenates on YPD agar. Each bar represents the mean + SD of 3 to 4 mice per group. Data are from one out of two independent experiments. (D,E) IL-10-deficient mice and WT controls were sublingually infected with C. albicans strain 101 and analyzed on day 9 post-infection. (D) Lymph node cells were re-stimulated and analyzed for IL-17 (left) and IFN-γ (right) production as in B. (E) Tongue fungal burdens were analyzed as in C. Each bar represents the mean + SD of 8–9 mice per group pooled from two independent experiments. Statistics were calculated using unpaired t-Test. *** p

    Article Snippet: Brefeldin A (10 μg/ml, AppliChem) was added for the last 5 h to inhibit the secretory pathway.

    Techniques: Mouse Assay, Infection, Flow Cytometry, Cytometry, Staining

    The Treg response during persistent colonization of the oral mucosa with  C. albicans .  (A–F)  WT mice were sublingually infected with strain 101 or SC5314 and cervical lymph node cells were analyzed on day 7 or day 18–21 as indicated.  (A,B)  Lymph node cells were re-stimulated with MutuDC1940 cells that were pulsed with heat-killed  C. albicans  or left unpulsed for 5 h in the presence of Brefeldin A. IL-17A production by CD3 + CD4 +  cells was analyzed by intracellular cytokine staining and flow cytometry.  (C,D)  The frequency of Foxp3-expressing cells within the CD4 +  lymphocyte compartment was assessed by flow cytometry.  (E,F)  PD-1, TIGIT, and Tim-3 expression by CD4 + Foxp3 +  Treg cells was analyzed by flow cytometry. ( G–J )  Il10 -Thy1.1 reporter mice were sublingually infected with strain 101 or SC5314 or left naive. IL-10 expression by Foxp3 +  Tregs and Foxp3 −  effector T cells was assessed in the cervical lymph nodes  (G,H)  and in the tongue  (I,J)  on day 21 post-infection by flow cytometry. Cells were pregated on CD90 + CD4 + (G,H)  or on CD45.2 + CD3 + (I,J) , respectively. Representative FACS plots are shown in (A, C, G, I); summary plots with data pooled from at least 2 experiments with 6–9 animals per infected group and 3–4 animals per naive group are shown in (B, D–F, H, J), with the exception of the right plot in  (D) , where data are from a single experiment with 3 animals per group. In B, Statistics were calculated using  t -test. In  (D–F, H, J) , statistics were calculated using one-way ANOVA. * p

    Journal: Frontiers in Immunology

    Article Title: Persistence of Candida albicans in the Oral Mucosa Induces a Curbed Inflammatory Host Response That Is Independent of Immunosuppression

    doi: 10.3389/fimmu.2019.00330

    Figure Lengend Snippet: The Treg response during persistent colonization of the oral mucosa with C. albicans . (A–F) WT mice were sublingually infected with strain 101 or SC5314 and cervical lymph node cells were analyzed on day 7 or day 18–21 as indicated. (A,B) Lymph node cells were re-stimulated with MutuDC1940 cells that were pulsed with heat-killed C. albicans or left unpulsed for 5 h in the presence of Brefeldin A. IL-17A production by CD3 + CD4 + cells was analyzed by intracellular cytokine staining and flow cytometry. (C,D) The frequency of Foxp3-expressing cells within the CD4 + lymphocyte compartment was assessed by flow cytometry. (E,F) PD-1, TIGIT, and Tim-3 expression by CD4 + Foxp3 + Treg cells was analyzed by flow cytometry. ( G–J ) Il10 -Thy1.1 reporter mice were sublingually infected with strain 101 or SC5314 or left naive. IL-10 expression by Foxp3 + Tregs and Foxp3 − effector T cells was assessed in the cervical lymph nodes (G,H) and in the tongue (I,J) on day 21 post-infection by flow cytometry. Cells were pregated on CD90 + CD4 + (G,H) or on CD45.2 + CD3 + (I,J) , respectively. Representative FACS plots are shown in (A, C, G, I); summary plots with data pooled from at least 2 experiments with 6–9 animals per infected group and 3–4 animals per naive group are shown in (B, D–F, H, J), with the exception of the right plot in (D) , where data are from a single experiment with 3 animals per group. In B, Statistics were calculated using t -test. In (D–F, H, J) , statistics were calculated using one-way ANOVA. * p

    Article Snippet: Brefeldin A (10 μg/ml, AppliChem) was added for the last 5 h to inhibit the secretory pathway.

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