recombinant il 2  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc recombinant il 2
    Representative flow cytometry showing presence of CD8 + T cell activation markers, a , PD-1 or b , CD25, following 4 day incubation with activation <t>cocktail</t> <t>(IL-2,</t> IL-15, anti-CD3, anti-CD28). c , Representative flow demonstrating cell surface PD-1 degradation on activated primary human CD8 + T cells following 24 hr treatment with 100 nM CXCL12-Nivo or nivolumab isotype.
    Recombinant Il 2, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant il 2/product/Gold Biotechnology Inc
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant il 2 - by Bioz Stars, 2024-05
    92/100 stars

    Images

    1) Product Images from "Modular cytokine receptor-targeting chimeras for targeted degradation of cell surface and extracellular proteins"

    Article Title: Modular cytokine receptor-targeting chimeras for targeted degradation of cell surface and extracellular proteins

    Journal: Nature Biotechnology

    doi: 10.1038/s41587-022-01456-2

    Representative flow cytometry showing presence of CD8 + T cell activation markers, a , PD-1 or b , CD25, following 4 day incubation with activation cocktail (IL-2, IL-15, anti-CD3, anti-CD28). c , Representative flow demonstrating cell surface PD-1 degradation on activated primary human CD8 + T cells following 24 hr treatment with 100 nM CXCL12-Nivo or nivolumab isotype.
    Figure Legend Snippet: Representative flow cytometry showing presence of CD8 + T cell activation markers, a , PD-1 or b , CD25, following 4 day incubation with activation cocktail (IL-2, IL-15, anti-CD3, anti-CD28). c , Representative flow demonstrating cell surface PD-1 degradation on activated primary human CD8 + T cells following 24 hr treatment with 100 nM CXCL12-Nivo or nivolumab isotype.

    Techniques Used: Flow Cytometry, Activation Assay, Incubation

    recombinant il 2  (Gold Biotechnology Inc)


    Bioz Verified Symbol Gold Biotechnology Inc is a verified supplier
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  • 92

    Structured Review

    Gold Biotechnology Inc recombinant il 2
    Representative flow cytometry showing presence of CD8 + T cell activation markers, a , PD-1 or b , CD25, following 4 day incubation with activation <t>cocktail</t> <t>(IL-2,</t> IL-15, anti-CD3, anti-CD28). c , Representative flow demonstrating cell surface PD-1 degradation on activated primary human CD8 + T cells following 24 hr treatment with 100 nM CXCL12-Nivo or nivolumab isotype.
    Recombinant Il 2, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant il 2/product/Gold Biotechnology Inc
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant il 2 - by Bioz Stars, 2024-05
    92/100 stars

    Images

    1) Product Images from "Modular cytokine receptor-targeting chimeras for targeted degradation of cell surface and extracellular proteins"

    Article Title: Modular cytokine receptor-targeting chimeras for targeted degradation of cell surface and extracellular proteins

    Journal: Nature Biotechnology

    doi: 10.1038/s41587-022-01456-2

    Representative flow cytometry showing presence of CD8 + T cell activation markers, a , PD-1 or b , CD25, following 4 day incubation with activation cocktail (IL-2, IL-15, anti-CD3, anti-CD28). c , Representative flow demonstrating cell surface PD-1 degradation on activated primary human CD8 + T cells following 24 hr treatment with 100 nM CXCL12-Nivo or nivolumab isotype.
    Figure Legend Snippet: Representative flow cytometry showing presence of CD8 + T cell activation markers, a , PD-1 or b , CD25, following 4 day incubation with activation cocktail (IL-2, IL-15, anti-CD3, anti-CD28). c , Representative flow demonstrating cell surface PD-1 degradation on activated primary human CD8 + T cells following 24 hr treatment with 100 nM CXCL12-Nivo or nivolumab isotype.

    Techniques Used: Flow Cytometry, Activation Assay, Incubation

    il2  (Gold Biotechnology Inc)


    Bioz Verified Symbol Gold Biotechnology Inc is a verified supplier
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    Structured Review

    Gold Biotechnology Inc il2
    (A-C) derive from CITE-seq data introduced in . (A) Volcano plot of top differentially expressed genes in CD301b + versus CD301b neg DC2 in D10 murine skin, complete list in Table 1 (B) Central functions enriched in each subset based on pathway analysis, complete list in Table 2 (C) Heat map of normalized average expression indicated genes in CD301b + versus CD301b neg cells in either CD11b hi or CD11b lo DC2 clusters and percentage of cells (circle size) in which expression was detected. All genes showed have a p-value <0.05 and were found in two independent scRNAseq experiments. (D-E) Spectral flow cytometry of activation markers in CD301b + versus CD301b neg DC2 from D10 skin 16h after colonization with S. epidermidis . (D) Representative histograms and gates used to delineate positive cells with corresponding (E) quantification of the % positive cells and geometrical mean of fluorescence intensity (gMFI) for the indicated markers. Each dot is biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (F) Schematic of the DC-T cell assay used in (G) and (H) to measure DC capacity to promote Tregs. CD301b + versus CD301b neg DC2 were separately sorted from the migDC fraction of D10 SDLN and incubated with OVA protein overnight then with CTV-labelled OT-II T cells in the presence <t>of</t> <t>IL-2</t> and TGF-β for 72h before flow cytometry to measure T cell proliferation. (G) Representative plots (pre-gated on Live CD4 + TCRβ + ). (H) Quantification of one of three representative experiments with technical replicates shown. One-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001.
    Il2, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il2/product/Gold Biotechnology Inc
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il2 - by Bioz Stars, 2024-05
    92/100 stars

    Images

    1) Product Images from "Early life tolerance depends on a subset of specialized dendritic cells and is reinforced by the skin microbiota"

    Article Title: Early life tolerance depends on a subset of specialized dendritic cells and is reinforced by the skin microbiota

    Journal: bioRxiv

    doi: 10.1101/2022.06.23.497363

    (A-C) derive from CITE-seq data introduced in . (A) Volcano plot of top differentially expressed genes in CD301b + versus CD301b neg DC2 in D10 murine skin, complete list in Table 1 (B) Central functions enriched in each subset based on pathway analysis, complete list in Table 2 (C) Heat map of normalized average expression indicated genes in CD301b + versus CD301b neg cells in either CD11b hi or CD11b lo DC2 clusters and percentage of cells (circle size) in which expression was detected. All genes showed have a p-value <0.05 and were found in two independent scRNAseq experiments. (D-E) Spectral flow cytometry of activation markers in CD301b + versus CD301b neg DC2 from D10 skin 16h after colonization with S. epidermidis . (D) Representative histograms and gates used to delineate positive cells with corresponding (E) quantification of the % positive cells and geometrical mean of fluorescence intensity (gMFI) for the indicated markers. Each dot is biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (F) Schematic of the DC-T cell assay used in (G) and (H) to measure DC capacity to promote Tregs. CD301b + versus CD301b neg DC2 were separately sorted from the migDC fraction of D10 SDLN and incubated with OVA protein overnight then with CTV-labelled OT-II T cells in the presence of IL-2 and TGF-β for 72h before flow cytometry to measure T cell proliferation. (G) Representative plots (pre-gated on Live CD4 + TCRβ + ). (H) Quantification of one of three representative experiments with technical replicates shown. One-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001.
    Figure Legend Snippet: (A-C) derive from CITE-seq data introduced in . (A) Volcano plot of top differentially expressed genes in CD301b + versus CD301b neg DC2 in D10 murine skin, complete list in Table 1 (B) Central functions enriched in each subset based on pathway analysis, complete list in Table 2 (C) Heat map of normalized average expression indicated genes in CD301b + versus CD301b neg cells in either CD11b hi or CD11b lo DC2 clusters and percentage of cells (circle size) in which expression was detected. All genes showed have a p-value <0.05 and were found in two independent scRNAseq experiments. (D-E) Spectral flow cytometry of activation markers in CD301b + versus CD301b neg DC2 from D10 skin 16h after colonization with S. epidermidis . (D) Representative histograms and gates used to delineate positive cells with corresponding (E) quantification of the % positive cells and geometrical mean of fluorescence intensity (gMFI) for the indicated markers. Each dot is biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (F) Schematic of the DC-T cell assay used in (G) and (H) to measure DC capacity to promote Tregs. CD301b + versus CD301b neg DC2 were separately sorted from the migDC fraction of D10 SDLN and incubated with OVA protein overnight then with CTV-labelled OT-II T cells in the presence of IL-2 and TGF-β for 72h before flow cytometry to measure T cell proliferation. (G) Representative plots (pre-gated on Live CD4 + TCRβ + ). (H) Quantification of one of three representative experiments with technical replicates shown. One-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001.

    Techniques Used: Expressing, Flow Cytometry, Activation Assay, Fluorescence, Incubation

    (A-B) D10 neonatal mice were colonized with S . epi -zsgreen and skin was harvested 16h later, with commensal antigen-loaded (zsgreen + ) and unloaded (zsgreen neg ) DCs separately sorts and submitted for scRNAseq. (A) UMAP of zsgreen + vs zsgreen neg DCs and (B) heatmaps comparing expression of select differentially genes between zsgreen + vs zsgreen neg CD301b + DC2 (CD301b status defined based Mgl2 expression). (C) Spectral flow cytometry comparing protein-level expression of key markers zsgreen + vs zsgreen neg CD301b + DC2 in neonatal skin 16h after colonization with S. epi- zsgreen colonization. Quantification of the % positive cells and gMFI for the indicated markers in loaded vs non loaded CD301b + DC2. Each dot is a biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (D-F) Human neonatal foreskin was incubated 4 h with S. epi -zsgreen as detailed in Fig. S3B and loaded (zsgreen + ) versus non-loaded (zsgreen neg ) live CD45 + CD16 neg HLADR + cells were separately sorted and submitted for scRNAseq. (D) UMAP of all cells combined, (E) UMAP of DC clusters split by zsgreen status, and (F) heat maps comparing expression of select genes between zsgreen + and zsgreen neg DC2 in human foreskin. (G) Schematic of the DC-T cell assay used in (H) and (I). Indicated DC subsets were sorted from the migDC fraction of D10 SDLN, incubated with S. epi -OVA overnight and then with CTV-labeled OT-II T cells for 72h in the presence of IL-2 and TGF-β. (H) Representative plots (pre-gated on Live CD4 + TCRβ + ) of showing percentage of OT-II Tregs and (I) and quantification thereof. Technical replicates are shown from one of 3 replicate experiments with similar results. One-Way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001, ns = not significant.
    Figure Legend Snippet: (A-B) D10 neonatal mice were colonized with S . epi -zsgreen and skin was harvested 16h later, with commensal antigen-loaded (zsgreen + ) and unloaded (zsgreen neg ) DCs separately sorts and submitted for scRNAseq. (A) UMAP of zsgreen + vs zsgreen neg DCs and (B) heatmaps comparing expression of select differentially genes between zsgreen + vs zsgreen neg CD301b + DC2 (CD301b status defined based Mgl2 expression). (C) Spectral flow cytometry comparing protein-level expression of key markers zsgreen + vs zsgreen neg CD301b + DC2 in neonatal skin 16h after colonization with S. epi- zsgreen colonization. Quantification of the % positive cells and gMFI for the indicated markers in loaded vs non loaded CD301b + DC2. Each dot is a biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (D-F) Human neonatal foreskin was incubated 4 h with S. epi -zsgreen as detailed in Fig. S3B and loaded (zsgreen + ) versus non-loaded (zsgreen neg ) live CD45 + CD16 neg HLADR + cells were separately sorted and submitted for scRNAseq. (D) UMAP of all cells combined, (E) UMAP of DC clusters split by zsgreen status, and (F) heat maps comparing expression of select genes between zsgreen + and zsgreen neg DC2 in human foreskin. (G) Schematic of the DC-T cell assay used in (H) and (I). Indicated DC subsets were sorted from the migDC fraction of D10 SDLN, incubated with S. epi -OVA overnight and then with CTV-labeled OT-II T cells for 72h in the presence of IL-2 and TGF-β. (H) Representative plots (pre-gated on Live CD4 + TCRβ + ) of showing percentage of OT-II Tregs and (I) and quantification thereof. Technical replicates are shown from one of 3 replicate experiments with similar results. One-Way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001, ns = not significant.

    Techniques Used: Expressing, Flow Cytometry, Incubation, Labeling

    murine il2 rmil2  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc murine il2 rmil2
    Murine Il2 Rmil2, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/murine il2 rmil2/product/Gold Biotechnology Inc
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    murine il2 rmil2 - by Bioz Stars, 2024-05
    92/100 stars

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    murine il2 rmil2  (Gold Biotechnology Inc)


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  • 92

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    Gold Biotechnology Inc murine il2 rmil2
    Murine Il2 Rmil2, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/murine il2 rmil2/product/Gold Biotechnology Inc
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    murine il2 rmil2 - by Bioz Stars, 2024-05
    92/100 stars

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    murine il2 rmil2  (Gold Biotechnology Inc)


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  • 92

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    Gold Biotechnology Inc murine il2 rmil2
    Murine Il2 Rmil2, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/murine il2 rmil2/product/Gold Biotechnology Inc
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    murine il2 rmil2 - by Bioz Stars, 2024-05
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    Gold Biotechnology Inc recombinant il 2
    Representative flow cytometry showing presence of CD8 + T cell activation markers, a , PD-1 or b , CD25, following 4 day incubation with activation <t>cocktail</t> <t>(IL-2,</t> IL-15, anti-CD3, anti-CD28). c , Representative flow demonstrating cell surface PD-1 degradation on activated primary human CD8 + T cells following 24 hr treatment with 100 nM CXCL12-Nivo or nivolumab isotype.
    Recombinant Il 2, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant il 2/product/Gold Biotechnology Inc
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant il 2 - by Bioz Stars, 2024-05
    92/100 stars
      Buy from Supplier

    92
    Gold Biotechnology Inc il2
    (A-C) derive from CITE-seq data introduced in . (A) Volcano plot of top differentially expressed genes in CD301b + versus CD301b neg DC2 in D10 murine skin, complete list in Table 1 (B) Central functions enriched in each subset based on pathway analysis, complete list in Table 2 (C) Heat map of normalized average expression indicated genes in CD301b + versus CD301b neg cells in either CD11b hi or CD11b lo DC2 clusters and percentage of cells (circle size) in which expression was detected. All genes showed have a p-value <0.05 and were found in two independent scRNAseq experiments. (D-E) Spectral flow cytometry of activation markers in CD301b + versus CD301b neg DC2 from D10 skin 16h after colonization with S. epidermidis . (D) Representative histograms and gates used to delineate positive cells with corresponding (E) quantification of the % positive cells and geometrical mean of fluorescence intensity (gMFI) for the indicated markers. Each dot is biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (F) Schematic of the DC-T cell assay used in (G) and (H) to measure DC capacity to promote Tregs. CD301b + versus CD301b neg DC2 were separately sorted from the migDC fraction of D10 SDLN and incubated with OVA protein overnight then with CTV-labelled OT-II T cells in the presence <t>of</t> <t>IL-2</t> and TGF-β for 72h before flow cytometry to measure T cell proliferation. (G) Representative plots (pre-gated on Live CD4 + TCRβ + ). (H) Quantification of one of three representative experiments with technical replicates shown. One-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001.
    Il2, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il2/product/Gold Biotechnology Inc
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il2 - by Bioz Stars, 2024-05
    92/100 stars
      Buy from Supplier

    92
    Gold Biotechnology Inc murine il2 rmil2
    (A-C) derive from CITE-seq data introduced in . (A) Volcano plot of top differentially expressed genes in CD301b + versus CD301b neg DC2 in D10 murine skin, complete list in Table 1 (B) Central functions enriched in each subset based on pathway analysis, complete list in Table 2 (C) Heat map of normalized average expression indicated genes in CD301b + versus CD301b neg cells in either CD11b hi or CD11b lo DC2 clusters and percentage of cells (circle size) in which expression was detected. All genes showed have a p-value <0.05 and were found in two independent scRNAseq experiments. (D-E) Spectral flow cytometry of activation markers in CD301b + versus CD301b neg DC2 from D10 skin 16h after colonization with S. epidermidis . (D) Representative histograms and gates used to delineate positive cells with corresponding (E) quantification of the % positive cells and geometrical mean of fluorescence intensity (gMFI) for the indicated markers. Each dot is biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (F) Schematic of the DC-T cell assay used in (G) and (H) to measure DC capacity to promote Tregs. CD301b + versus CD301b neg DC2 were separately sorted from the migDC fraction of D10 SDLN and incubated with OVA protein overnight then with CTV-labelled OT-II T cells in the presence <t>of</t> <t>IL-2</t> and TGF-β for 72h before flow cytometry to measure T cell proliferation. (G) Representative plots (pre-gated on Live CD4 + TCRβ + ). (H) Quantification of one of three representative experiments with technical replicates shown. One-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001.
    Murine Il2 Rmil2, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/murine il2 rmil2/product/Gold Biotechnology Inc
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    murine il2 rmil2 - by Bioz Stars, 2024-05
    92/100 stars
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    Image Search Results


    Representative flow cytometry showing presence of CD8 + T cell activation markers, a , PD-1 or b , CD25, following 4 day incubation with activation cocktail (IL-2, IL-15, anti-CD3, anti-CD28). c , Representative flow demonstrating cell surface PD-1 degradation on activated primary human CD8 + T cells following 24 hr treatment with 100 nM CXCL12-Nivo or nivolumab isotype.

    Journal: Nature Biotechnology

    Article Title: Modular cytokine receptor-targeting chimeras for targeted degradation of cell surface and extracellular proteins

    doi: 10.1038/s41587-022-01456-2

    Figure Lengend Snippet: Representative flow cytometry showing presence of CD8 + T cell activation markers, a , PD-1 or b , CD25, following 4 day incubation with activation cocktail (IL-2, IL-15, anti-CD3, anti-CD28). c , Representative flow demonstrating cell surface PD-1 degradation on activated primary human CD8 + T cells following 24 hr treatment with 100 nM CXCL12-Nivo or nivolumab isotype.

    Article Snippet: Following CD8 + T cell isolation, cells were stimulated with recombinant IL-2 (GoldBio) and IL-15 (GoldBio) and ImmunoCult Human CD3/CD28 T cell Activation (STEMCELL Technologies) for 4 d at 37 °C.

    Techniques: Flow Cytometry, Activation Assay, Incubation

    (A-C) derive from CITE-seq data introduced in . (A) Volcano plot of top differentially expressed genes in CD301b + versus CD301b neg DC2 in D10 murine skin, complete list in Table 1 (B) Central functions enriched in each subset based on pathway analysis, complete list in Table 2 (C) Heat map of normalized average expression indicated genes in CD301b + versus CD301b neg cells in either CD11b hi or CD11b lo DC2 clusters and percentage of cells (circle size) in which expression was detected. All genes showed have a p-value <0.05 and were found in two independent scRNAseq experiments. (D-E) Spectral flow cytometry of activation markers in CD301b + versus CD301b neg DC2 from D10 skin 16h after colonization with S. epidermidis . (D) Representative histograms and gates used to delineate positive cells with corresponding (E) quantification of the % positive cells and geometrical mean of fluorescence intensity (gMFI) for the indicated markers. Each dot is biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (F) Schematic of the DC-T cell assay used in (G) and (H) to measure DC capacity to promote Tregs. CD301b + versus CD301b neg DC2 were separately sorted from the migDC fraction of D10 SDLN and incubated with OVA protein overnight then with CTV-labelled OT-II T cells in the presence of IL-2 and TGF-β for 72h before flow cytometry to measure T cell proliferation. (G) Representative plots (pre-gated on Live CD4 + TCRβ + ). (H) Quantification of one of three representative experiments with technical replicates shown. One-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001.

    Journal: bioRxiv

    Article Title: Early life tolerance depends on a subset of specialized dendritic cells and is reinforced by the skin microbiota

    doi: 10.1101/2022.06.23.497363

    Figure Lengend Snippet: (A-C) derive from CITE-seq data introduced in . (A) Volcano plot of top differentially expressed genes in CD301b + versus CD301b neg DC2 in D10 murine skin, complete list in Table 1 (B) Central functions enriched in each subset based on pathway analysis, complete list in Table 2 (C) Heat map of normalized average expression indicated genes in CD301b + versus CD301b neg cells in either CD11b hi or CD11b lo DC2 clusters and percentage of cells (circle size) in which expression was detected. All genes showed have a p-value <0.05 and were found in two independent scRNAseq experiments. (D-E) Spectral flow cytometry of activation markers in CD301b + versus CD301b neg DC2 from D10 skin 16h after colonization with S. epidermidis . (D) Representative histograms and gates used to delineate positive cells with corresponding (E) quantification of the % positive cells and geometrical mean of fluorescence intensity (gMFI) for the indicated markers. Each dot is biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (F) Schematic of the DC-T cell assay used in (G) and (H) to measure DC capacity to promote Tregs. CD301b + versus CD301b neg DC2 were separately sorted from the migDC fraction of D10 SDLN and incubated with OVA protein overnight then with CTV-labelled OT-II T cells in the presence of IL-2 and TGF-β for 72h before flow cytometry to measure T cell proliferation. (G) Representative plots (pre-gated on Live CD4 + TCRβ + ). (H) Quantification of one of three representative experiments with technical replicates shown. One-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001.

    Article Snippet: For suboptimal Treg inducing conditions, IL2 (200 U/mL, GoldBio, # 1310-02-100) and TGFβ (2 ng/mL, Peprotech, #100-21C-10UG) were added to the wells.

    Techniques: Expressing, Flow Cytometry, Activation Assay, Fluorescence, Incubation

    (A-B) D10 neonatal mice were colonized with S . epi -zsgreen and skin was harvested 16h later, with commensal antigen-loaded (zsgreen + ) and unloaded (zsgreen neg ) DCs separately sorts and submitted for scRNAseq. (A) UMAP of zsgreen + vs zsgreen neg DCs and (B) heatmaps comparing expression of select differentially genes between zsgreen + vs zsgreen neg CD301b + DC2 (CD301b status defined based Mgl2 expression). (C) Spectral flow cytometry comparing protein-level expression of key markers zsgreen + vs zsgreen neg CD301b + DC2 in neonatal skin 16h after colonization with S. epi- zsgreen colonization. Quantification of the % positive cells and gMFI for the indicated markers in loaded vs non loaded CD301b + DC2. Each dot is a biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (D-F) Human neonatal foreskin was incubated 4 h with S. epi -zsgreen as detailed in Fig. S3B and loaded (zsgreen + ) versus non-loaded (zsgreen neg ) live CD45 + CD16 neg HLADR + cells were separately sorted and submitted for scRNAseq. (D) UMAP of all cells combined, (E) UMAP of DC clusters split by zsgreen status, and (F) heat maps comparing expression of select genes between zsgreen + and zsgreen neg DC2 in human foreskin. (G) Schematic of the DC-T cell assay used in (H) and (I). Indicated DC subsets were sorted from the migDC fraction of D10 SDLN, incubated with S. epi -OVA overnight and then with CTV-labeled OT-II T cells for 72h in the presence of IL-2 and TGF-β. (H) Representative plots (pre-gated on Live CD4 + TCRβ + ) of showing percentage of OT-II Tregs and (I) and quantification thereof. Technical replicates are shown from one of 3 replicate experiments with similar results. One-Way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001, ns = not significant.

    Journal: bioRxiv

    Article Title: Early life tolerance depends on a subset of specialized dendritic cells and is reinforced by the skin microbiota

    doi: 10.1101/2022.06.23.497363

    Figure Lengend Snippet: (A-B) D10 neonatal mice were colonized with S . epi -zsgreen and skin was harvested 16h later, with commensal antigen-loaded (zsgreen + ) and unloaded (zsgreen neg ) DCs separately sorts and submitted for scRNAseq. (A) UMAP of zsgreen + vs zsgreen neg DCs and (B) heatmaps comparing expression of select differentially genes between zsgreen + vs zsgreen neg CD301b + DC2 (CD301b status defined based Mgl2 expression). (C) Spectral flow cytometry comparing protein-level expression of key markers zsgreen + vs zsgreen neg CD301b + DC2 in neonatal skin 16h after colonization with S. epi- zsgreen colonization. Quantification of the % positive cells and gMFI for the indicated markers in loaded vs non loaded CD301b + DC2. Each dot is a biological replicate. 1 of 2 independent experiments are shown. Two-way ANOVA with Šídák’s multiple comparisons test. (D-F) Human neonatal foreskin was incubated 4 h with S. epi -zsgreen as detailed in Fig. S3B and loaded (zsgreen + ) versus non-loaded (zsgreen neg ) live CD45 + CD16 neg HLADR + cells were separately sorted and submitted for scRNAseq. (D) UMAP of all cells combined, (E) UMAP of DC clusters split by zsgreen status, and (F) heat maps comparing expression of select genes between zsgreen + and zsgreen neg DC2 in human foreskin. (G) Schematic of the DC-T cell assay used in (H) and (I). Indicated DC subsets were sorted from the migDC fraction of D10 SDLN, incubated with S. epi -OVA overnight and then with CTV-labeled OT-II T cells for 72h in the presence of IL-2 and TGF-β. (H) Representative plots (pre-gated on Live CD4 + TCRβ + ) of showing percentage of OT-II Tregs and (I) and quantification thereof. Technical replicates are shown from one of 3 replicate experiments with similar results. One-Way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001, ns = not significant.

    Article Snippet: For suboptimal Treg inducing conditions, IL2 (200 U/mL, GoldBio, # 1310-02-100) and TGFβ (2 ng/mL, Peprotech, #100-21C-10UG) were added to the wells.

    Techniques: Expressing, Flow Cytometry, Incubation, Labeling