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il 6  (R&D Systems)


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    R&D Systems il 6
    a, Violin plots showing expression of Prg4 , Thy1 and Cd34 across fibroblast clusters. b, Heatmap of top 20 DEGs defining lining fibroblasts (LFs : Prg4 ⁺) and five sublining fibroblast populations (SLFs : Myoc ⁺, Ugdh ⁺, Cxcl12 ⁺, Col3a1 ⁺ and Ccl11 ⁺). c, Violin plots showing expression of representative marker genes across fibroblast subtypes. d, UMAPs showing fibroblast clusters in young and aged mice, with relative cluster proportions by age group. e, Gene Ontology enrichment analysis of genes upregulated in young versus aged fibroblast clusters. Dot size indicates gene ratio and color indicates adjusted P value. f, CellChat analysis <t>of</t> <t>IL-6</t> signaling networks in young and aged synovium, showing inferred communication probabilities among cell populations. g, SenMayo module scores across fibroblast clusters.
    Il 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 97/100, based on 1096 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+il+6+duoset+elisa/Mouse+IL-6+DuoSet+ELISA/bio_rxiv__64898__2026__04__17__719179-148-6-8
    Average 97 stars, based on 1096 article reviews
    il 6 - by Bioz Stars, 2026-10
    97/100 stars

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    1) Product Images from "Single-cell atlas reveals age-related cellular shifts underlying fibrosis in murine synovium"

    Article Title: Single-cell atlas reveals age-related cellular shifts underlying fibrosis in murine synovium

    Journal: bioRxiv

    doi: 10.64898/2026.04.17.719179

    a, Violin plots showing expression of Prg4 , Thy1 and Cd34 across fibroblast clusters. b, Heatmap of top 20 DEGs defining lining fibroblasts (LFs : Prg4 ⁺) and five sublining fibroblast populations (SLFs : Myoc ⁺, Ugdh ⁺, Cxcl12 ⁺, Col3a1 ⁺ and Ccl11 ⁺). c, Violin plots showing expression of representative marker genes across fibroblast subtypes. d, UMAPs showing fibroblast clusters in young and aged mice, with relative cluster proportions by age group. e, Gene Ontology enrichment analysis of genes upregulated in young versus aged fibroblast clusters. Dot size indicates gene ratio and color indicates adjusted P value. f, CellChat analysis of IL-6 signaling networks in young and aged synovium, showing inferred communication probabilities among cell populations. g, SenMayo module scores across fibroblast clusters.
    Figure Legend Snippet: a, Violin plots showing expression of Prg4 , Thy1 and Cd34 across fibroblast clusters. b, Heatmap of top 20 DEGs defining lining fibroblasts (LFs : Prg4 ⁺) and five sublining fibroblast populations (SLFs : Myoc ⁺, Ugdh ⁺, Cxcl12 ⁺, Col3a1 ⁺ and Ccl11 ⁺). c, Violin plots showing expression of representative marker genes across fibroblast subtypes. d, UMAPs showing fibroblast clusters in young and aged mice, with relative cluster proportions by age group. e, Gene Ontology enrichment analysis of genes upregulated in young versus aged fibroblast clusters. Dot size indicates gene ratio and color indicates adjusted P value. f, CellChat analysis of IL-6 signaling networks in young and aged synovium, showing inferred communication probabilities among cell populations. g, SenMayo module scores across fibroblast clusters.

    Techniques Used: Expressing, Marker

    a , Serum IL-6 levels in young and aged male mice (n = 8 per group). b , Representative flow cytometry gating strategy for peripheral blood myeloid cells. Bar graphs show the quantification of Ly6G + neutrophils, Ly6C high monocytes and Ly6C low monocytes per ml of blood in young and aged male mice (n = 9 per group). c , Representative flow cytometry gating strategy for synovial tissue monocytes and neutrophils. Flow cytometry-based enumeration of Ly6C + monocytes and Ly6G + neutrophils per paw in young and aged male mice (n = 6 per group). Data are mean ± s.e.m. Two-tailed Mann-Whitney U test. * P < 0.05, *** P < 0.001; ns, not significant.
    Figure Legend Snippet: a , Serum IL-6 levels in young and aged male mice (n = 8 per group). b , Representative flow cytometry gating strategy for peripheral blood myeloid cells. Bar graphs show the quantification of Ly6G + neutrophils, Ly6C high monocytes and Ly6C low monocytes per ml of blood in young and aged male mice (n = 9 per group). c , Representative flow cytometry gating strategy for synovial tissue monocytes and neutrophils. Flow cytometry-based enumeration of Ly6C + monocytes and Ly6G + neutrophils per paw in young and aged male mice (n = 6 per group). Data are mean ± s.e.m. Two-tailed Mann-Whitney U test. * P < 0.05, *** P < 0.001; ns, not significant.

    Techniques Used: Flow Cytometry, Two Tailed Test, MANN-WHITNEY

    a , Heatmap showing expression of top 20 genes across synovial macrophage clusters, identifying Pf4 + , Vsig4 + and Cd74 + macrophage subsets. b , Violin plots depict expression of representative marker genes across the three macrophage subsets. c , Gene Ontology enrichment analysis of genes upregulated in young versus aged macrophages within each subset. Dot size indicates gene ratio and color indicates adjusted P value. d , IL-6 and TNF-α secretion by FACS-sorted synovial macrophages from young and aged mice following 24 h ex vivo stimulation with or without LPS (n = 10 mice per group; one-way ANOVA test). e , Phagocytosis assay showing representative pHrodo Green fluorescence histograms and frequency of pHrodo + cells within CD64 + F4/80 + synovial macrophages, 2 h after intra-articular injection of pHrodo Green E. coli BioParticles (n = 6 mice per group; two-tailed Mann-Whitney U test). FMO control is shown in grey. Data are mean ± s.e.m. ** P < 0.01, **** P < 0.0001.
    Figure Legend Snippet: a , Heatmap showing expression of top 20 genes across synovial macrophage clusters, identifying Pf4 + , Vsig4 + and Cd74 + macrophage subsets. b , Violin plots depict expression of representative marker genes across the three macrophage subsets. c , Gene Ontology enrichment analysis of genes upregulated in young versus aged macrophages within each subset. Dot size indicates gene ratio and color indicates adjusted P value. d , IL-6 and TNF-α secretion by FACS-sorted synovial macrophages from young and aged mice following 24 h ex vivo stimulation with or without LPS (n = 10 mice per group; one-way ANOVA test). e , Phagocytosis assay showing representative pHrodo Green fluorescence histograms and frequency of pHrodo + cells within CD64 + F4/80 + synovial macrophages, 2 h after intra-articular injection of pHrodo Green E. coli BioParticles (n = 6 mice per group; two-tailed Mann-Whitney U test). FMO control is shown in grey. Data are mean ± s.e.m. ** P < 0.01, **** P < 0.0001.

    Techniques Used: Expressing, Marker, Ex Vivo, Phagocytosis Assay, Fluorescence, Injection, Two Tailed Test, MANN-WHITNEY, Control

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    Article Title: Montelukast suppresses NLRP3 inflammasome activation as a potential prophylactic agent against gout arthritis flares
    Article Snippet: WT BMDM supernatants from the NLRC4/AIM2 inflammasome assays were evaluated for IL-1β using the Mouse IL-1β/IL-1F2 DuoSet ELISA (R&D Systems, Cat. No. DY401) following manufacturer’s instructions.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Montelukast suppresses NLRP3 inflammasome activation as a potential prophylactic agent against gout arthritis flares.
    Article Snippet: WT BMDM supernatants from the NLRC4/AIM2 inflammasome assays were evaluated for IL1β using the Mouse IL-1β/IL-1F2 DuoSet ELISA (R&D Systems, Cat. No. DY401) following manufacturer’s instructions. .. WT BMDM supernatants from the montelukast inflammation assays were evaluated for IL-6 using the Mouse IL-6 DuoSet ELISA (R&D Systems, Cat. No. DY406), TNF-α using the Mouse TNF-α DuoSet ELISA (R&D Systems, Cat. No. DY410), and CXCL1 using the Mouse CXCL1/KC DuoSet ELISA (R&D Systems, Cat. No. DY453) following manufacturer’s instructions. .. AR TIC LE IN PR ES S (R&D Systems, Cat. No. DY401), IL-18 using the Mouse IL-18 DuoSet ELISA (R&D Systems, Cat. No. DY7625), IL-6 using the Mouse IL-6 DuoSet ELISA (R&D Systems, Cat. No. DY406), CXCL1 using the Mouse CXCL1/KC DuoSet ELISA (R&D Systems, Cat. No. DY453), and TNF-α using the Mouse TNF-α DuoSet ELISA (R&D Systems, Cat. No. DY410) following manufacturer’s instructions.

    Article Title: SIRT7 inhibits cerebral ischemic injury by inhibiting microglia M1 polarization via desuccinylation of NAMPT.
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    Article Title: Montelukast suppresses NLRP3 inflammasome activation as a potential prophylactic agent against gout arthritis flares
    Article Snippet: WT BMDM supernatants from the NLRC4/AIM2 inflammasome assays were evaluated for IL-1β using the Mouse IL-1β/IL-1F2 DuoSet ELISA (R&D Systems, Cat. No. DY401) following manufacturer’s instructions. .. WT BMDM supernatants from the montelukast inflammation assays were evaluated for IL-6 using the Mouse IL-6 DuoSet ELISA (R&D Systems, Cat. No. DY406), TNF-α using the Mouse TNF-α DuoSet ELISA (R&D Systems, Cat. No. DY410), and CXCL1 using the Mouse CXCL1/KC DuoSet ELISA (R&D Systems, Cat. No. DY453) following manufacturer’s instructions. .. Mouse paw homogenates were assayed for IL-1β using the Mouse IL-1β/IL-1F2 DuoSet ELISA (R&D Systems, Cat. No. DY401), IL-18 using the Mouse IL-18 DuoSet ELISA (R&D Systems, Cat. No. DY7625), IL-6 using the Mouse IL-6 DuoSet ELISA (R&D Systems, Cat. No. DY406), CXCL1 using the Mouse CXCL1/KC DuoSet ELISA (R&D Systems, Cat. No. DY453), and TNF-α using the Mouse TNF-α DuoSet ELISA (R&D Systems, Cat. No. DY410) following manufacturer’s instructions.

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    Article Snippet: The amount of mitochondrial DNA in each fraction was analyzed by qPCR using PowerUP SYBR Green PCR Master Mix (Applied Biosystems) with a Step One Real-Time PCR System (Applied Biosystems). .. For IL-1β measurements, cells were preincubated with 20 ng/ml of LPS for 3 h and stimulated with 50 μg/ml IMQ, 50 μg/ml RSQ, 5 mM ATP, or 5 μM nigericin for 30 min or 3 h. In the case of pDCs, the cells were seeded onto 96-well plates at a density of 2.0 × 10 cells per well and stimulated with ligands for 24 h. The cytokine levels of IL-6, TNF-α, and IL-1β in the culture supernatant were measured using a mouse IL-6 DuoSet ELISA (R&D Systems), mouse TNF-α ELISA kit (Invitrogen), and mouse IL-1β ELISA kit (Invitrogen), respectively, according to the manufacturer’s instructions. .. For IL-1β measurements, cells were preincubated with 20 ng/ml of LPS for 3 h and stimulated with 50 μg/ml IMQ, 50 μg/ml RSQ, 5 mM ATP, or 5 μM nigericin for 30 min or 3 h. In the case of pDCs, the cells were seeded onto 96-well plates at a density of 2.0 × 10 cells per well and stimulated with ligands for 24 h. The cytokine levels of IL-6, TNF-α, and IL-1β in the culture supernatant were measured using a mouse IL-6 DuoSet ELISA (R&D Systems), mouse TNF-α ELISA kit (Invitrogen), and mouse IL-1β ELISA kit (Invitrogen), respectively, according to the manufacturer’s instructions.

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    Sandwich ELISA:

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    Image Search Results


    a, Violin plots showing expression of Prg4 , Thy1 and Cd34 across fibroblast clusters. b, Heatmap of top 20 DEGs defining lining fibroblasts (LFs : Prg4 ⁺) and five sublining fibroblast populations (SLFs : Myoc ⁺, Ugdh ⁺, Cxcl12 ⁺, Col3a1 ⁺ and Ccl11 ⁺). c, Violin plots showing expression of representative marker genes across fibroblast subtypes. d, UMAPs showing fibroblast clusters in young and aged mice, with relative cluster proportions by age group. e, Gene Ontology enrichment analysis of genes upregulated in young versus aged fibroblast clusters. Dot size indicates gene ratio and color indicates adjusted P value. f, CellChat analysis of IL-6 signaling networks in young and aged synovium, showing inferred communication probabilities among cell populations. g, SenMayo module scores across fibroblast clusters.

    Journal: bioRxiv

    Article Title: Single-cell atlas reveals age-related cellular shifts underlying fibrosis in murine synovium

    doi: 10.64898/2026.04.17.719179

    Figure Lengend Snippet: a, Violin plots showing expression of Prg4 , Thy1 and Cd34 across fibroblast clusters. b, Heatmap of top 20 DEGs defining lining fibroblasts (LFs : Prg4 ⁺) and five sublining fibroblast populations (SLFs : Myoc ⁺, Ugdh ⁺, Cxcl12 ⁺, Col3a1 ⁺ and Ccl11 ⁺). c, Violin plots showing expression of representative marker genes across fibroblast subtypes. d, UMAPs showing fibroblast clusters in young and aged mice, with relative cluster proportions by age group. e, Gene Ontology enrichment analysis of genes upregulated in young versus aged fibroblast clusters. Dot size indicates gene ratio and color indicates adjusted P value. f, CellChat analysis of IL-6 signaling networks in young and aged synovium, showing inferred communication probabilities among cell populations. g, SenMayo module scores across fibroblast clusters.

    Article Snippet: Mouse TNFα (DY410, R&D Systems) and IL-6 (DY406, R&D Systems) concentrations in serum samples and cell culture supernatants were measured by ELISA according to the manufacturer’s instructions.

    Techniques: Expressing, Marker

    a , Serum IL-6 levels in young and aged male mice (n = 8 per group). b , Representative flow cytometry gating strategy for peripheral blood myeloid cells. Bar graphs show the quantification of Ly6G + neutrophils, Ly6C high monocytes and Ly6C low monocytes per ml of blood in young and aged male mice (n = 9 per group). c , Representative flow cytometry gating strategy for synovial tissue monocytes and neutrophils. Flow cytometry-based enumeration of Ly6C + monocytes and Ly6G + neutrophils per paw in young and aged male mice (n = 6 per group). Data are mean ± s.e.m. Two-tailed Mann-Whitney U test. * P < 0.05, *** P < 0.001; ns, not significant.

    Journal: bioRxiv

    Article Title: Single-cell atlas reveals age-related cellular shifts underlying fibrosis in murine synovium

    doi: 10.64898/2026.04.17.719179

    Figure Lengend Snippet: a , Serum IL-6 levels in young and aged male mice (n = 8 per group). b , Representative flow cytometry gating strategy for peripheral blood myeloid cells. Bar graphs show the quantification of Ly6G + neutrophils, Ly6C high monocytes and Ly6C low monocytes per ml of blood in young and aged male mice (n = 9 per group). c , Representative flow cytometry gating strategy for synovial tissue monocytes and neutrophils. Flow cytometry-based enumeration of Ly6C + monocytes and Ly6G + neutrophils per paw in young and aged male mice (n = 6 per group). Data are mean ± s.e.m. Two-tailed Mann-Whitney U test. * P < 0.05, *** P < 0.001; ns, not significant.

    Article Snippet: Mouse TNFα (DY410, R&D Systems) and IL-6 (DY406, R&D Systems) concentrations in serum samples and cell culture supernatants were measured by ELISA according to the manufacturer’s instructions.

    Techniques: Flow Cytometry, Two Tailed Test, MANN-WHITNEY

    a , Heatmap showing expression of top 20 genes across synovial macrophage clusters, identifying Pf4 + , Vsig4 + and Cd74 + macrophage subsets. b , Violin plots depict expression of representative marker genes across the three macrophage subsets. c , Gene Ontology enrichment analysis of genes upregulated in young versus aged macrophages within each subset. Dot size indicates gene ratio and color indicates adjusted P value. d , IL-6 and TNF-α secretion by FACS-sorted synovial macrophages from young and aged mice following 24 h ex vivo stimulation with or without LPS (n = 10 mice per group; one-way ANOVA test). e , Phagocytosis assay showing representative pHrodo Green fluorescence histograms and frequency of pHrodo + cells within CD64 + F4/80 + synovial macrophages, 2 h after intra-articular injection of pHrodo Green E. coli BioParticles (n = 6 mice per group; two-tailed Mann-Whitney U test). FMO control is shown in grey. Data are mean ± s.e.m. ** P < 0.01, **** P < 0.0001.

    Journal: bioRxiv

    Article Title: Single-cell atlas reveals age-related cellular shifts underlying fibrosis in murine synovium

    doi: 10.64898/2026.04.17.719179

    Figure Lengend Snippet: a , Heatmap showing expression of top 20 genes across synovial macrophage clusters, identifying Pf4 + , Vsig4 + and Cd74 + macrophage subsets. b , Violin plots depict expression of representative marker genes across the three macrophage subsets. c , Gene Ontology enrichment analysis of genes upregulated in young versus aged macrophages within each subset. Dot size indicates gene ratio and color indicates adjusted P value. d , IL-6 and TNF-α secretion by FACS-sorted synovial macrophages from young and aged mice following 24 h ex vivo stimulation with or without LPS (n = 10 mice per group; one-way ANOVA test). e , Phagocytosis assay showing representative pHrodo Green fluorescence histograms and frequency of pHrodo + cells within CD64 + F4/80 + synovial macrophages, 2 h after intra-articular injection of pHrodo Green E. coli BioParticles (n = 6 mice per group; two-tailed Mann-Whitney U test). FMO control is shown in grey. Data are mean ± s.e.m. ** P < 0.01, **** P < 0.0001.

    Article Snippet: Mouse TNFα (DY410, R&D Systems) and IL-6 (DY406, R&D Systems) concentrations in serum samples and cell culture supernatants were measured by ELISA according to the manufacturer’s instructions.

    Techniques: Expressing, Marker, Ex Vivo, Phagocytosis Assay, Fluorescence, Injection, Two Tailed Test, MANN-WHITNEY, Control

    (A) Illustration of experimental outline ( https://biorender.com/ ). (B) Principal component analysis of gene expression patterns comparing different combinations of primed (P) and licensed (L) CD8 + T cells by cDCs or MoDCs (n= 4). (C) Hierarchal cluster of significantly changed genes in primed (P) and licensed (L) CD8 + T cells (T) by cDCs or MoDCs (n= 4). (D) Circus plots showing connecting unique MoDC ligands with receptors on antigen-experienced T cells (n=4). (E)B16F10 tumor cell counts in IncuCyte following incubation with effector CD8 + T cells and sup fraction of LPS activated MoDC. (F) Volcano plot showing differential expression of proteins between 6-hour LPS-activated MoDCs and Naive MoDCs (>30 k Da fraction) (n=3). (G) GSEA enrichment plot for the “TNF Signalling Pathway,” showing the distribution of leading-edge genes along the ranked gene list. The plot highlights the correlation between gene rank and pathway enrichment, with the running enrichment score (ES) shown on top. The vertical line marks the position of the leading edge (n=3). (H) Proteomap (level 3) showing enriched signalling pathways from significantly and non-significantly upregulated proteins in activated MoDCs (>30 kDa fraction) (n=3). (I) IncuCyte live-cell counts of B16F10 melanoma cells over time following incubation with OVA-primed CD8 + T cells with or without the addition of TNF. Shown is the summary of one experiment out of at least three indecent experiments. (J) ELISA analysis of TNF-alpha cytokine productions by naïve, LPS or CL307 actiavted MoDCs vs classical DCs, differentiated from bone marrow and the spleen of C57BL/6 mice, respectively (n=3). Statistical significance was calculated using ANOVA with Tukey correction for multiple comparisons. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

    Journal: bioRxiv

    Article Title: Successful dendritic cell vaccines require lasting in-situ TNFa secretion to license antitumor CD8 + T cell cytotoxicity

    doi: 10.64898/2026.04.06.716539

    Figure Lengend Snippet: (A) Illustration of experimental outline ( https://biorender.com/ ). (B) Principal component analysis of gene expression patterns comparing different combinations of primed (P) and licensed (L) CD8 + T cells by cDCs or MoDCs (n= 4). (C) Hierarchal cluster of significantly changed genes in primed (P) and licensed (L) CD8 + T cells (T) by cDCs or MoDCs (n= 4). (D) Circus plots showing connecting unique MoDC ligands with receptors on antigen-experienced T cells (n=4). (E)B16F10 tumor cell counts in IncuCyte following incubation with effector CD8 + T cells and sup fraction of LPS activated MoDC. (F) Volcano plot showing differential expression of proteins between 6-hour LPS-activated MoDCs and Naive MoDCs (>30 k Da fraction) (n=3). (G) GSEA enrichment plot for the “TNF Signalling Pathway,” showing the distribution of leading-edge genes along the ranked gene list. The plot highlights the correlation between gene rank and pathway enrichment, with the running enrichment score (ES) shown on top. The vertical line marks the position of the leading edge (n=3). (H) Proteomap (level 3) showing enriched signalling pathways from significantly and non-significantly upregulated proteins in activated MoDCs (>30 kDa fraction) (n=3). (I) IncuCyte live-cell counts of B16F10 melanoma cells over time following incubation with OVA-primed CD8 + T cells with or without the addition of TNF. Shown is the summary of one experiment out of at least three indecent experiments. (J) ELISA analysis of TNF-alpha cytokine productions by naïve, LPS or CL307 actiavted MoDCs vs classical DCs, differentiated from bone marrow and the spleen of C57BL/6 mice, respectively (n=3). Statistical significance was calculated using ANOVA with Tukey correction for multiple comparisons. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

    Article Snippet: Levels of secreted TNF-alpha cytokine in 24h LPS-activated, 48h after wash and LPS/CL307 reactivated supernatants of mouse/human MoDCs were quantified using a sandwich ELISA kit (R&D Systems, Cat. No. DY406).

    Techniques: Gene Expression, Incubation, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay

    (A) ELISA analysis of TNF-alpha cytokine productions by cells isolated from the tumor of tumor bearing mice on different days (n=6). (B) Mean percentage of CD45.1 MoDCs detected in B16F10 tumors 48 hours after vaccination. MoDCs were activated overnight with LPS, pulsed with gp100 peptide, and injected either s.c. or i.v. into CD45.2⁺ mice 10 days after tumor challenge (n=3 or 4). (C) FACS analysis of CD45.1 DCs expressing TNF in vitro and CD45.1 DCs isolated from the tumor of B16F10 tumor-bearing C75BL/6 mice (day 10/11; n = 5). (D) Mean percentage of CD86 and TNF expression on CD45.1⁺ MoDCs detected in B16F10 tumors at 24- and 72-hours post-vaccination. MoDCs were activated overnight with LPS, pulsed with gp100 peptide, and injected s.c. into CD45.2 mice 10 days after B16F10 tumor challenge (n=2,3,4 or 5). (E) Expression of CD86/CD40 activation markers on MoDCs differentiated from the bone barrow of C57BL/6 mice, after LPS or PolyIC activation and wash. (n=3). (F) IncuCyte live-cell counts of B16F10 melanoma cells over time following incubation with primed CD8+ T cells with or without LPS-activated or washed MoDCs sup (n=3). (G) ELISA analysis of TNF-alpha cytokine productions by mouse MoDCs differentiated from bone marrow of gp100 mice, and underwent LPS treatment, wash and LPS/CL307 reactivation (n=3).. (H) ELISA analysis of TNF-alpha cytokine productions by mouse MoDCs differentiated from bone marrow of gp100 mice, and underwent LPS+INFγ treatment, wash and LPS/CL307 reactivation. (I) Expression of CD86/CD40 activation markers on MoDCs differentiated from the bone barrow of C57BL/6 mice, after LPS/PolyIC activation, wash and reactivation (n=3). (J) IncuCyte live-cell counts of H2b-tdt gp100 loaded B16 melanoma cells over time following incubation with naïve or primed CD8 cells with or without LPS-activated or washed MoDCs sup (n=3). (K) ELISA analysis of TNF-alpha cytokine productions by MoDCs differentiated from blood monocytes of healthy donors, and underwent LPS treatment, wash and reactivation. (L) IncuCyte live-cell counts of A375 human melanoma cells over time following incubation with 1G4 TCR infected CD8 T cells with or without LPS-activated or washed human MoDCs sup. Shown is the summary of one experiment out of at least three indecent experiments. Statistical significance was calculated using Two-way ANOVA with Tukey correction for multiple comparisons. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

    Journal: bioRxiv

    Article Title: Successful dendritic cell vaccines require lasting in-situ TNFa secretion to license antitumor CD8 + T cell cytotoxicity

    doi: 10.64898/2026.04.06.716539

    Figure Lengend Snippet: (A) ELISA analysis of TNF-alpha cytokine productions by cells isolated from the tumor of tumor bearing mice on different days (n=6). (B) Mean percentage of CD45.1 MoDCs detected in B16F10 tumors 48 hours after vaccination. MoDCs were activated overnight with LPS, pulsed with gp100 peptide, and injected either s.c. or i.v. into CD45.2⁺ mice 10 days after tumor challenge (n=3 or 4). (C) FACS analysis of CD45.1 DCs expressing TNF in vitro and CD45.1 DCs isolated from the tumor of B16F10 tumor-bearing C75BL/6 mice (day 10/11; n = 5). (D) Mean percentage of CD86 and TNF expression on CD45.1⁺ MoDCs detected in B16F10 tumors at 24- and 72-hours post-vaccination. MoDCs were activated overnight with LPS, pulsed with gp100 peptide, and injected s.c. into CD45.2 mice 10 days after B16F10 tumor challenge (n=2,3,4 or 5). (E) Expression of CD86/CD40 activation markers on MoDCs differentiated from the bone barrow of C57BL/6 mice, after LPS or PolyIC activation and wash. (n=3). (F) IncuCyte live-cell counts of B16F10 melanoma cells over time following incubation with primed CD8+ T cells with or without LPS-activated or washed MoDCs sup (n=3). (G) ELISA analysis of TNF-alpha cytokine productions by mouse MoDCs differentiated from bone marrow of gp100 mice, and underwent LPS treatment, wash and LPS/CL307 reactivation (n=3).. (H) ELISA analysis of TNF-alpha cytokine productions by mouse MoDCs differentiated from bone marrow of gp100 mice, and underwent LPS+INFγ treatment, wash and LPS/CL307 reactivation. (I) Expression of CD86/CD40 activation markers on MoDCs differentiated from the bone barrow of C57BL/6 mice, after LPS/PolyIC activation, wash and reactivation (n=3). (J) IncuCyte live-cell counts of H2b-tdt gp100 loaded B16 melanoma cells over time following incubation with naïve or primed CD8 cells with or without LPS-activated or washed MoDCs sup (n=3). (K) ELISA analysis of TNF-alpha cytokine productions by MoDCs differentiated from blood monocytes of healthy donors, and underwent LPS treatment, wash and reactivation. (L) IncuCyte live-cell counts of A375 human melanoma cells over time following incubation with 1G4 TCR infected CD8 T cells with or without LPS-activated or washed human MoDCs sup. Shown is the summary of one experiment out of at least three indecent experiments. Statistical significance was calculated using Two-way ANOVA with Tukey correction for multiple comparisons. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

    Article Snippet: Levels of secreted TNF-alpha cytokine in 24h LPS-activated, 48h after wash and LPS/CL307 reactivated supernatants of mouse/human MoDCs were quantified using a sandwich ELISA kit (R&D Systems, Cat. No. DY406).

    Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Injection, Expressing, In Vitro, Activation Assay, Incubation, Infection