human cytokine array panel a  (R&D Systems)

Journal: Aging Cell

Article Title: Targeting Hyperoxia‐Induced Cellular Senescence in Developing Human Airway Cells: Senomorphics Versus Senolytics Versus Antioxidants

doi: 10.1111/acel.70538

Figure Lengend Snippet: Hyperoxia‐induced senescent fASM secrete higher levels of SASP. Cells cultured till day 7 in normoxic (21% O 2 ) or hyperoxic (50% O 2 ) environment were incubated with fresh growth media in normoxia for 24 h, and the supernatants were collected. Samples were then analyzed for SASP secretion by Eve Technologies Corporation (Calgary, Alberta, Canada) using Luminex xMAP and the Human Cytokine/Chemokine 96‐Plex Discovery Assay Array (HD96) panel. (A) Out of the 96 proteins, 80 were detected and visualized using a heatmap. (B) A detailed analysis of each detected marker is shown as fold‐change normalized to control. For statistical analysis, 2‐way ANOVA with a two‐stage linear step‐up procedure of Benjamini, Krieger and Yekutieli test, with individual variances computed for each comparison was applied. False discovery rate < 0.05 was considered significant. Data are presented as box plot of n = 7 cell lines per group.

Article Snippet: Supernatants collected at day 8 from normoxia and hyperoxia‐exposed fASM were analyzed for SASP secretion by Eve Technologies Corporation (Calgary, Alberta, Canada) using the Human Cytokine/Chemokine 96‐Plex Discovery Assay Array (HD96) panel.

Techniques: Cell Culture, Incubation, Luminex, Marker, Control, Comparison

Journal: Prostate International

Article Title: Interleukin-6 secreted by tumor-associated macrophages promotes proliferation and migration through JAK2/STAT3 signaling pathway in human prostate cancer cells

doi: 10.1016/j.prnil.2025.11.002

Figure Lengend Snippet: Morphological characteristics of THP-1-derived macrophages after cytokine or PC3-CM stimulation. THP-1 monocytes were differentiated into unpolarized macrophages (M0) using PMA treatment for 24 h. M0 macrophages were subsequently polarized into M1 macrophages by stimulation with LPS and IFN-γ, into M2 macrophages by stimulation with IL-4 and IL-13, or into TAMs using conditioned medium from PC3 prostate cancer cells. Microscopy images show that M0 macrophages displayed a round and less-elongated morphology, whereas M1 macrophages exhibited elongated spindle-like shapes with prominent protrusions (yellow arrowheads). M2 macrophages also developed elongated morphologies but with fewer extensions compared to M1. TAMs demonstrated heterogeneous morphologies resembling M2-like phenotypes, characterized by mixed elongated and irregularly spread cells (yellow arrowheads). Images were captured after 48 h of polarization. (magnification: upper × 200, bottom × 400).

Article Snippet: Human cytokine array panel A (R&D Systems) was incubated with blocking buffer and then with the sample-antibody mixture overnight at 4°C.

Techniques: Derivative Assay, Microscopy

Journal: Prostate International

Article Title: Interleukin-6 secreted by tumor-associated macrophages promotes proliferation and migration through JAK2/STAT3 signaling pathway in human prostate cancer cells

doi: 10.1016/j.prnil.2025.11.002

Figure Lengend Snippet: Cytokine secretion profiles of macrophages, PC3 cells, and their co-culture. (A) Cytokine array analysis of conditioned media collected from M0 macrophages, TAMs, PC3 prostate cancer cells, and TAMs co-cultured with PC3 cells for 48 h. Black dots indicate the presence of specific cytokines, each of which corresponds to a name listed in the table below. (B) Quantitative analysis of dot blot intensity revealed that TAMs and TAM + PC3 conditions induced higher levels of proinflammatory and tumor-promoting cytokines, including CCL2/MCP-1, MIP-1α, CXCL1/GROα, IL-1β, IL-6, IL-8, TNF-α, G-CSF, GM-CSF, and MIF, compared with M0 or PC3 cells alone. Data are presented as mean ± SEM. ∗ P < 0.05 versus M0. IL, interleukin; MIT, migration inhibitory factor; TAMs, tumor-associated macrophages.

Article Snippet: Human cytokine array panel A (R&D Systems) was incubated with blocking buffer and then with the sample-antibody mixture overnight at 4°C.

Techniques: Co-Culture Assay, Cell Culture, Quantitative Dot Blot, Migration

Journal: International Journal of Molecular Sciences

Article Title: HO-1197 as a Multifaceted Therapeutic: Targeting the Cell Cycle, Angiogenesis, Metastasis, and Tumor Immunity in Hepatocellular Carcinoma

doi: 10.3390/ijms262110329

Figure Lengend Snippet: HO-1197 promotes pro-inflammatory cytokine production in the TME. ( A ) THP-1 cells were treated with HO-1197 (starting at 10 mg/mL) using 2-fold serial dilutions for 48 h. Cell survival was calculated by counting nuclei. ( B ) The levels of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-8) secreted from THP-1 cells treated with 0, 0.313, or 1.25 mg/mL HO-1197 were detected using ELISAs. ( C – E ) A cytokine array was performed (upper), and cytokine levels were quantified (lower) in Huh7 ( C ) and Hep3B ( D ) cells treated with HO-1197 (0 or 1 mg/mL) and in Huh7 cells treated with HO-1089 (0 or 1 mg/mL) ( E ). Data are expressed as the mean ± SD (n = 3). ** p < 0.005 and *** p < 0.0005 compared with the control group.

Article Snippet: A Human Cytokine Array (R&D Systems) was used to quantify pro-inflammatory cytokines in cell lysates, according to the manufacturer’s protocol.

Techniques: Control

Journal: Frontiers in Medicine

Article Title: Pseudomonas aeruginosa elastase down-regulates host inflammatory responses by degrading cytokines and chemokines: a non-healing wound perspective

doi: 10.3389/fmed.2025.1585252

Figure Lengend Snippet: Elastase-containing conditioned medium reduces cytokine and chemokine levels. Human monocytes (A) or 25% human whole blood (B) were incubated for 18–22 h at 37°C with 0.1–10% conditioned medium of P. aeruginosa strains PAO1, PAOB1 or clinical isolate 15159 and supernatants were analyzed using ELISA. Results are means ± SEM of 6–9 (A) or 10–13 (B) experiments. Values are significantly (* p < 0.05, ** p < 0.005, *** p < 0.0005) different as analyzed using a repeated measures one-way ANOVA with a Tukey’s multiple comparisons test. (C) Heatmap of normalized cytokine and chemokine (both CXC and CC) levels obtained with Proteome Profiler™ Arrays of whole blood stimulated with 10% CM or non-stimulated cells as a control ( n = 4) (See also ).

Article Snippet: Cytokine levels in human whole blood and monocyte culture samples were assessed using BioSource CytoSetTM (Invitrogen) and the Proteome ProfilerTM Human Cytokine Array Panel A (R&D systems) following manufacturer’s instructions.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Medicine

Article Title: Pseudomonas aeruginosa elastase down-regulates host inflammatory responses by degrading cytokines and chemokines: a non-healing wound perspective

doi: 10.3389/fmed.2025.1585252

Figure Lengend Snippet: P. aeruginosa elastase reduces cytokine and chemokine levels. (A) Human monocytes were incubated for 18–22 h at 37°C with LPS (100 ng/mL) and a range of P. aeruginosa elastase (ELA). Results are means ± SEM of 3 experiments. Values are significantly (* p < 0.05, ** p < 0.005, *** p < 0.0005) different compared to the control as analyzed using a repeated measures one-way ANOVA with a Dunnett’s multiple comparisons test. (B) Heatmap of normalized cytokine and chemokine (CXC and CC sub-families) levels obtained with Proteome Profiler™ Arrays of 25% human whole blood cultures stimulated with LPS (100 ng/mL) and elastase (2 units/mL; n = 4) (see also ).

Article Snippet: Cytokine levels in human whole blood and monocyte culture samples were assessed using BioSource CytoSetTM (Invitrogen) and the Proteome ProfilerTM Human Cytokine Array Panel A (R&D systems) following manufacturer’s instructions.

Techniques: Incubation, Control

Journal: Frontiers in Medicine

Article Title: Pseudomonas aeruginosa elastase down-regulates host inflammatory responses by degrading cytokines and chemokines: a non-healing wound perspective

doi: 10.3389/fmed.2025.1585252

Figure Lengend Snippet: P. aeruginosa elastase does not inhibit intracellular cytokine production. Human whole blood supplemented with 2 μM monensin was incubated for 5 h with (A,B) 10% CM or (C) LPS (1 μg/mL) with or without elastase (2 units/mL) and intracellular cytokine levels were measured using FACS. Results are the average median fluorescent intensities (MFI) ± SEM of 3 (C) and 4 (A , two batches of CM per donor ) blood donors. Values are significantly (* p < 0.05, ** p < 0.005) different as analyzed using a repeated measures one-way ANOVA with a Tukey’s multiple comparisons test. (B) Representative examples of the FACS plots: dot plots of forward-sideward scatter and gating of monocytes based on CD14 expression, and histograms of CD14 expression of the three cell types and of the five tested cytokines in monocytes.

Article Snippet: Cytokine levels in human whole blood and monocyte culture samples were assessed using BioSource CytoSetTM (Invitrogen) and the Proteome ProfilerTM Human Cytokine Array Panel A (R&D systems) following manufacturer’s instructions.

Techniques: Incubation, Expressing

Journal: Frontiers in Medicine

Article Title: Pseudomonas aeruginosa elastase down-regulates host inflammatory responses by degrading cytokines and chemokines: a non-healing wound perspective

doi: 10.3389/fmed.2025.1585252

Figure Lengend Snippet: P. aeruginosa elastase degrades cytokines and chemokines. Plasma derived from mice injected with LPS was incubated with a range of elastase for 2 h (A) and 24 h (B) , and cytokine levels were analyzed using FACS. Results are normalized values of 4 independent experiments and are significantly (* p < 0.05, ** p < 0.005, *** p < 0.0005) different from the control as analyzed using a repeated measures one-way ANOVA with a Dunnett’s multiple comparisons test. (C) Recombinant cytokines and (D) chemokines were incubated with elastase (1 U/mL) for 1, 3 or 24 h and degradation was visualized using SDS-PAGE. C1, control incubated for 1 h; C24, control incubated for 24 h. (E) Effect of 2 and 4 units elastase/mL on IL-8 and MCP-1 after 24 h incubation. (F) To investigate whether cytokines and chemokines are degraded in infected wounds, wound fluids from non-infected acute wounds (aWF) or P. aeruginosa colonized/infected venous leg ulcers (VLU) were spiked with recombinant human TNF-α or G-CSF and incubated for 24 h at 37°C, followed by analysis using SDS-PAGE and western blotting (P, plasma used as a control; representative blots are shown).

Article Snippet: Cytokine levels in human whole blood and monocyte culture samples were assessed using BioSource CytoSetTM (Invitrogen) and the Proteome ProfilerTM Human Cytokine Array Panel A (R&D systems) following manufacturer’s instructions.

Techniques: Clinical Proteomics, Derivative Assay, Injection, Incubation, Control, Recombinant, SDS Page, Infection, Western Blot

Journal: Foods

Article Title: Structural Elucidation of Heteropolysaccharides from the Peach-Shaped Dictyophora indusiata and Its Anti-Inflammatory Activity

doi: 10.3390/foods14091536

Figure Lengend Snippet: Effects of DIVP on the human cytokines and phosphokinase. ( A , B ) Dot blot array analysis ( A ) and quantification ( B ) of pro-inflammatory cytokine secretion from LPS-stimulated U937-macrophages in the presence or absence of 0.2 mg/mL DIVP. ( C , D ) Dot blot array analysis ( C ) and relative quantification ( D ) of human phosphokinase from LPS-stimulated U937 macrophages in the presence or absence of 0.2 mg/mL DIVP. A, B, C, D, E in Fig. 6A represent different rows. Statistical significance was determined with a one-way or two-way ANOVA (for dot blot assay), and p values are as follows: ns, not significant; * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. LPS group.

Article Snippet: The levels of soluble cytokines secreted by LPS-stimulated U937 cells, both in the presence and absence of DIVP, were analyzed using the Human Cytokine Array Kit panel A (ARY005B, R&D Systems, Minneapolis, MN, USA) following the manufacturer’s instructions.

Techniques: Dot Blot, Quantitative Proteomics, Control