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Enzo Life Sciences cd40l
Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min <t>CD40L</t> stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
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1) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

2) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

3) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

4) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

5) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

6) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

7) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

8) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

9) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

10) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

11) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

12) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

13) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

14) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

15) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

16) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

17) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

18) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

19) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

20) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

21) Product Images from "Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response"

Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

doi: 10.1002/cyto.a.23737

Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
Figure Legend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

Techniques Used: Expressing, Mass Cytometry

CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p
Figure Legend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

Techniques Used: Fluorescence, Flow Cytometry

CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p
Figure Legend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

Techniques Used: Binding Assay, Fluorescence, Flow Cytometry

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    Enzo Life Sciences cd40l
    Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min <t>CD40L</t> stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p
    Cd40l, supplied by Enzo Life Sciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

    Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

    Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

    doi: 10.1002/cyto.a.23737

    Figure Lengend Snippet: Reduced CD40-mediated NFκB p65 phosphorylation in GC B cells despite increased surface CD40 expression NFκB p65 phosphorylation (p-p65) at serine 529 following 15 min CD40L stimulation was measured by mass cytometry. Populations were gated on biaxial plots; GC B cells as CD3 − CD19 + CD20 ++ CD38 + and non-GC B cells as CD3 − CD19 + CD38 − . The non-GC population was further gated to enrich for IgD + naïve B cells (possibly containing unswitched memory B cells) and IgD − memory B cells. A) Histograms of p-p65 from one representative tonsil (T19). B-C) Relative induction of p-p65 by CD40L (B) and surface CD40 expression in unstimulated cells (C) was measured simultaneously. Mean ± SD. * p

    Article Snippet: Despite differences in total p-p65, comparable signaling kinetics were observed; p-p65 levels peaked at 15 minutes following CD40L in all three B-cell subsets.

    Techniques: Expressing, Mass Cytometry

    CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

    Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

    Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

    doi: 10.1002/cyto.a.23737

    Figure Lengend Snippet: CD40L signaling responses distinguish GC B cells from naïve and memory B cells in human tonsils. CD40L-induced signaling (A: total IκBα, B: p-p65-S529, C: p-S6-Ser235/236) in human tonsillar cells was measured by fluorescence flow cytometry. Values represent MFI relative to unstimulated naïve B cells in one representative tonsil (A-C) or mean values ± SEM (D-F), n = 3–4 individual tonsils. After debarcoding, GC B cells were gated as CD20 hi CD38 + , non-GC (CD20 + CD38 − ) were split into CD27 − IgD + naïve and CD27 + IgD − memory B cells. * p

    Article Snippet: Despite differences in total p-p65, comparable signaling kinetics were observed; p-p65 levels peaked at 15 minutes following CD40L in all three B-cell subsets.

    Techniques: Fluorescence, Flow Cytometry

    CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

    Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

    Article Title: Human germinal center B cells differ from naïve and memory B cells in CD40 expression and CD40L-induced signaling response

    doi: 10.1002/cyto.a.23737

    Figure Lengend Snippet: CD40L-induced IκBα degradation is not proportional to level of CD40L binding CD40L-induced signaling (IκBα and p-p65-S529) in human tonsillar cells was measured by fluorescence flow cytometry as a function of ligand uptake (bound CD40L). Cells were stimulated with CD40L and a secondary antibody was used to detect bound CD40L after permeabilization of the cells. Non-GC (naïve and memory) B cells were gated as CD20 + CD38 − and GC B cells as CD20 hi CD38 + . (A) The cells were further gated into 7 populations based on the level of bound CD40L. (B-C) Signaling is shown as arcsinh ratio relative to the lowest level of bound CD40L in non-GC B cells. D) Signaling responses is shown as arcsinh ratio relative to the unstimulated condition for each cell type and for each level of CD40L. (C-D) Mean values ± SEM, n = 4 individual tonsils. * p

    Article Snippet: Despite differences in total p-p65, comparable signaling kinetics were observed; p-p65 levels peaked at 15 minutes following CD40L in all three B-cell subsets.

    Techniques: Binding Assay, Fluorescence, Flow Cytometry