elisa kit  (R&D Systems)

Journal: Physiological Reports

Article Title: Sex differences in IL ‐6 responses to moderate‐intensity aerobic exercise are explained by cycling power output

doi: 10.14814/phy2.70850

Figure Lengend Snippet: Unadjusted exercise‐induced changes in IL‐6 in males and females (Model 1). Estimated marginal means of IL‐6 derived from linear mixed‐effects Model 1 (without covariates) were back‐transformed from the log scale using the exponential function and are presented in the original units. Error bars represent 95% confidence intervals. These differences became nonsignificant in Model 2, in which mean power output during exercise was included as a covariate. Sample size: MP and LP, n = 12. LP, luteal phase; MP, menstrual phase.

Article Snippet: IL‐6 concentrations were measured in serum using a human IL‐6 ELISA kit (Quantikine HS600C, R&D Systems, Minneapolis, MN, USA). d‐ROMs were measured in serum using a commercial kit (d‐ROMs Test Kit, DI‐003b, Wismerll Co., Ltd., Tokyo, Japan).

Techniques: Derivative Assay, Transformation Assay

Journal: Communications Biology

Article Title: Epigenome-wide association study of circulating interleukin-6 connects DNA methylation to immunometabolic and inflammatory health

doi: 10.1038/s42003-026-09520-2

Figure Lengend Snippet: a Scatterplot comparing effect sizes from the base model (unadjusted) versus a model adjusted for twelve predicted blood cell-type proportions. CpG sites with insufficient evidence for an independent IL-6 effect ( p fdr ≥ 0.05) after adjustment are highlighted in yellow ( n = 130). b Scatterplot comparing base model effect sizes versus a model adjusted for smoking status. CpGs previously removed due to cell-type confounding are shown in grey and one CpG removed due to smoking effects is highlighted in yellow. Data underlying Fig. 2 is provided in Supplementary Data 02 .

Article Snippet: Serum concentrations of IL-6 were measured with the Human IL-6 Quantikine HS ELISA kit (R&D Systems, Wiesbaden, Germany), and hsCRP was determined by a high-sensitivity latex-enhanced nephelometric assay on a BN II analyzer (Dade Behring) .

Techniques:

Journal: Communications Biology

Article Title: Epigenome-wide association study of circulating interleukin-6 connects DNA methylation to immunometabolic and inflammatory health

doi: 10.1038/s42003-026-09520-2

Figure Lengend Snippet: a Directional Manhattan plot showing the genomic position of all 412,226 tested CpGs ( x -axis) and their signed log 10 p -value ( y -axis). CpGs positively associated with IL-6 are displayed in the upper half, and inversely correlated CpGs in the lower half. The 401 CpGs associated with IL-6 after multiple testing correction and sensitivity analyses are highlighted in blue (odd chromosomes) and yellow (even chromosomes), while grey dots represent all other tested CpGs. b Effect sizes and 95% confidence intervals (CIs) for the 401 IL-6-associated CpGs, with points coloured by mean DNAm level. c Volcano plot showing the effect size ( x -axis) against −log 10 p -value ( y -axis) for all tested CpGs. Significant CpGs are coloured by mean DNAm level and the top hits are labelled, while all other CpGs are shown in grey. Data underlying Fig. 3 is provided in Supplementary Data 01 .

Article Snippet: Serum concentrations of IL-6 were measured with the Human IL-6 Quantikine HS ELISA kit (R&D Systems, Wiesbaden, Germany), and hsCRP was determined by a high-sensitivity latex-enhanced nephelometric assay on a BN II analyzer (Dade Behring) .

Techniques:

Journal: Communications Biology

Article Title: Epigenome-wide association study of circulating interleukin-6 connects DNA methylation to immunometabolic and inflammatory health

doi: 10.1038/s42003-026-09520-2

Figure Lengend Snippet: a Scatterplot comparing IL-6 associated DNAm effect sizes before and after adjustment for hsCRP levels. b Scatterplot comparing hsCRP-associated DNAm effect sizes before and after adjustment for IL-6 levels. The Pearson correlation coefficient (R) and associated p -value are shown in blue for both plots. Data underlying Fig. 4 is provided in Supplementary Data 01, 03, and 04 .

Article Snippet: Serum concentrations of IL-6 were measured with the Human IL-6 Quantikine HS ELISA kit (R&D Systems, Wiesbaden, Germany), and hsCRP was determined by a high-sensitivity latex-enhanced nephelometric assay on a BN II analyzer (Dade Behring) .

Techniques:

Journal: Communications Biology

Article Title: Epigenome-wide association study of circulating interleukin-6 connects DNA methylation to immunometabolic and inflammatory health

doi: 10.1038/s42003-026-09520-2

Figure Lengend Snippet: a Forest plot of ORs and 95% CIs with enrichment across 14 chromatin states, calculated using the Roadmap PBMC reference epigenome (E062) with all tested CpGs as the background set. States are sorted by OR, with blue denoting enrichment, yellow indicating depletion, and grey for non-significant results. One non-significant term (ZNF/Rpts) not shown due to an extremely wide 95% CI. b Bar plot of −log 10 p -values from TFBS enrichment analysis, performed using HOMER on sequences within 50 bp of IL-6 associated CpGs, tested against a GC-matched random genomic background. Data underlying Fig. 6 is provided in Supplementary Data 06 and 08 .

Article Snippet: Serum concentrations of IL-6 were measured with the Human IL-6 Quantikine HS ELISA kit (R&D Systems, Wiesbaden, Germany), and hsCRP was determined by a high-sensitivity latex-enhanced nephelometric assay on a BN II analyzer (Dade Behring) .

Techniques:

Journal: Communications Biology

Article Title: Epigenome-wide association study of circulating interleukin-6 connects DNA methylation to immunometabolic and inflammatory health

doi: 10.1038/s42003-026-09520-2

Figure Lengend Snippet: a Top five enriched gene sets, plus eleven enriched terms with specific relevance to Th17 activation and IL-6 signalling. Bars are colored by the percentage of overlapping genes; genes mapped onto the aetiological cascade from T-cell activation to glycolysis-fuelled Th17 proliferation are indicated. b Top ten transcription factors (TFs) whose regulons are most enriched in the genes linked to IL-6-associated DNAm using DoRothEA TF target annotations. c Selected enriched terms (white boxes) and mapped genes (black boxes) arranged along the pathway to Th17 differentiation, including four activation signals: (i) TCR activation, (ii) co-stimulation, (iii) IL-1β stimulation, (iv) IL-6/JAK/STAT3 signalling, culminating in (v) mTORC1-driven glycolysis fuelling rapid proliferation and effector functions. Created in BioRender. Data underlying Fig. 7 is provided in Supplementary Data and .

Article Snippet: Serum concentrations of IL-6 were measured with the Human IL-6 Quantikine HS ELISA kit (R&D Systems, Wiesbaden, Germany), and hsCRP was determined by a high-sensitivity latex-enhanced nephelometric assay on a BN II analyzer (Dade Behring) .

Techniques: Activation Assay

Journal: Communications Biology

Article Title: Epigenome-wide association study of circulating interleukin-6 connects DNA methylation to immunometabolic and inflammatory health

doi: 10.1038/s42003-026-09520-2

Figure Lengend Snippet: a Scatter plot comparing the observed effect (PGS-DNAm associations) with the predicted effect (combining PGS effect on IL-6 and IL-6 CpG associations). Correlation coefficients (R) and associated p -values are shown in blue. b Scatter plot comparing the observed (mQTL IL-6 associations) and predicted effects (combined effect of SNP on DNAm, and DNAm on IL-6). Each point represents an IL-6-associated CpG. Data underlying Fig. 8 is provided in Supplementary Data and .

Article Snippet: Serum concentrations of IL-6 were measured with the Human IL-6 Quantikine HS ELISA kit (R&D Systems, Wiesbaden, Germany), and hsCRP was determined by a high-sensitivity latex-enhanced nephelometric assay on a BN II analyzer (Dade Behring) .

Techniques:

Journal: Communications Biology

Article Title: Epigenome-wide association study of circulating interleukin-6 connects DNA methylation to immunometabolic and inflammatory health

doi: 10.1038/s42003-026-09520-2

Figure Lengend Snippet: a Forest plot showing the 52 genes linked to IL-6-associated DNAm that were responsive to IL-6 stimulation in isolated immune cells in vitro. b Forest plots of CpGs ranked in the top ten for at least one tested trait in the mediation analysis. Bars are coloured by the direction and significance of the mediated effect: orange for significant positive mediation, blue for significant negative mediation, and grey for non-significant findings after correcting for multiple testing. Data underlying Fig. is provided in Supplementary Data and .

Article Snippet: Serum concentrations of IL-6 were measured with the Human IL-6 Quantikine HS ELISA kit (R&D Systems, Wiesbaden, Germany), and hsCRP was determined by a high-sensitivity latex-enhanced nephelometric assay on a BN II analyzer (Dade Behring) .

Techniques: Isolation, In Vitro

Journal: Advanced Science

Article Title: CD3ɛ Nanobody‐Engineered Extracellular Vesicles Driving In Vivo Generation of TCE‐secreting CAR‐Ts for Solid Tumor Therapy With Memory Response and Minimal Immunogenicity

doi: 10.1002/advs.202519440

Figure Lengend Snippet: CD3ε‐Nb EVs showed higher transfection efficiency with minimal immunogenicity risk than lipid and viral‐carrier‐based approaches. (A) Schematic protocol for evaluating the immunological responses of CD3ε‐Nb mEVs, liposomes, LNPs, and LVs in an immunocompetent mouse model. C57BL/6 mice ( n = 5) were treated with or without 2 × 10 9 particles of CD3ε Nb‐conjugated liposomes, LNPs, NIH/3T3‐derived CD3ε Nb‐mouse CD63‐expressing EVs (CD3ε‐Nb mEVs), or CD3ε‐Nb LVs weekly for four weeks through tail vein injection. All mice were sacrificed, and their serum and splenocytes were collected. (B) The serum contents of C1q, ALT, IFN‐α, IL‐6, IFN‐γ, and IL‐1β were detected using ELISA kits according to the user instructions. (C) The presence of IgM and IgG against CD3ε Nb‐expressing liposomes, LNPs, LVs, or mEVs in the serum was determined by an ELISA‐based assay through coating with CD3ε‐Nb liposomes, LNPs, LVs, or mEVs. (D) The Nb‐CAR‐expressing splenocytes were determined by flow cytometry using specific antibodies against VHH and mouse CD3. (E–J) Minimal influence on EV‐mediated in viv o Nb‐CAR.TCE‐T generation by prior exposure of lipid‐based carriers. (E) The expression of HLA‐G on HLA‐G‐overexpressing GL261‐luc cells was measured by flow cytometry. (F) Representative protocol for investigating the effect of prior exposure with liposomes and LNPs on subsequent transgene delivery using CD3ε‐Nb mEVs in a tumor‐bearing syngeneic mouse model. C57BL/6 mice ( n = 3 and 5) were orthotopically implanted with GL261 cells (2 × 10 5 cells) and subsequently treated with 2 × 10 9 particles of CD3ε Nb‐conjugated liposomes, LNPs, NIH/3T3‐derived CD3ε Nb‐mouse CD63‐expressing EVs (CD3ε‐Nb mEVs), or CD3ε‐Nb LVs weekly for two weeks through tail vein injection. The mice were then infused with CD3ε‐Nb liposomes, LNPs, or mEVs loaded with the Nb‐CAR.TCE transgene (1 mg/kg) in the following two weeks. (G) All mice were sacrificed on day 21 after the first infusion, and the expression of Nb‐CAR on splenocytes was measured by flow cytometry using VHH and mouse CD3‐specific antibodies (G). Or (H–J) the tumor growth was monitored weekly using the IVIS system to detect the bioluminescent signals (H,I), and the survival rates of these mice were documented (J). The in vivo tumor growth data were presented as mean ± SEM and analyzed using one‐way ANOVA to compare means across multiple groups. The survival rate was analyzed using the Kaplan–Meier method and log‐rank test. Statistical significance was set at p ‐value < 0.05. * p < 0.05, ** p < 0.01, *** p < 0.001. The results of the experiments were analyzed using one‐way ANOVA to compare means across multiple groups. Data are mean ± SD, * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The levels of human IL‐6 (#HS600C), mouse IL‐6 (#M6000B), human IFN‐γ (#DIF50C), mouse C1q (#MCD930), mouse IL‐6, (#MFNAS0), mouse IFN‐α (#MIF00), mouse IFN‐γ (#MIF00), human TNF‐α (#DTA00D), mouse TNF‐α (#MTA00B), human IL‐1β (#HSLB00D), and mouse IL‐1β (#MHSLB00) were measured, along with the multiplex cytokine array set (which includes human CCL3/MIP‐1α, CCL4/MIP‐1β, CD25/IL‐2Rα, GM‐CSF, IFN‐γ, IL‐1β, IL‐2, IL‐4, IL‐5, IL‐6, IL‐8/CXCL8, IL‐10, IL‐12/IL‐23 p40, IL‐17/IL‐17A, and TNF‐α), in the serum samples and coculture supernatants using ELISA kits (R&D Systems) and ABplex Human Cytokine 15‐Plex Assay Kit (RK05202, Abclonal), following the manufacturer's instructions.

Techniques: Transfection, Immunopeptidomics, Liposomes, Derivative Assay, Expressing, Injection, Enzyme-linked Immunosorbent Assay, Flow Cytometry, In Vivo