quantikine elisa  (R&D Systems)

Journal: Stem Cell Research & Therapy

Article Title: IL-1β modulates inflammatory response of human bone marrow-derived MSCs and neutrophil recruitment in vitro via NF-kB-associated signaling

doi: 10.1186/s13287-026-05029-x

Figure Lengend Snippet: IL-1β stimulation increases expression of neutrophil recruitment and NF-kB signaling genes by BM-hMSCs. Heatmap clustering of the Z-score of the top 20 differentially expressed genes between unstimulated control BM-hMSCs (3 replicates, 1 donor, green) and IL-1 β stimulated BM-hMSCs (3 replicates, 1 donor, purple). Red color indicates higher expression of the genes and blue color indicates decreased expression A . Protein expression of CXCL1 (10 replicates, 4 donors), CCL2 (4 replicates, 2 donors), CXCL5 (4 replicates, 2 donors), and CXCL8/IL-8 (7 replicates, 3 donors) in BM-hMSCs secretome measured by ELLA or ELISA. Data are presented as median, and statistical analysis was performed using unpaired t-test with Welch’s correction B . Bar plot of the top 20 significant Gene Ontology (GO) biological processes representing IL- 1β induced upregulated differentially expressed genes C IL-1 β, Interleukin-1β; BM-hMSCs, Bone marrow derived human mesenchymal cells; CCL2, chemokine (C–C motif) ligand 2; CXCL1, chemokine (C-X-C motif) ligand 1; CXCL2, chemokine (C-X-C motif) ligand 1; CXCL8, chemokine (C-X-C motif) ligand 8; ****, p < 0,0001

Article Snippet: CXCL1 was measured in diluted 1:1 or undiluted CM samples using Quantikine ELISA (cat# DGR00B, R&D Systems).

Techniques: Expressing, Control, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: Radiation Oncology (London, England)

Article Title: CXCR2 affects sensitization of radioresistant HPV-negative head and neck squamous cell carcinoma cells by ABT-263

doi: 10.1186/s13014-026-02798-w

Figure Lengend Snippet: ABT-263 decreases senescence and alters the SASP in UPCI:SCC040 and Cal33 cells. Cells were treated with 1 µM ABT-263 (Cal33) or 5 µM of ABT-263 (UPCI:SCC040) concomitantly with 6 Gy irradiation and analyzed on D4 or D6. ( A-B ) Gene expression of secreted factors measured by qRT-PCR and normalized to unirradiated DMSO controls. ( A ) UPCI:SCC040. ( B ) Cal33. ( C-D ) Secreted IL1A, IL1B, IL8 and CXCL1 protein levels were detected by ELISA on D4 and D6. Protein concentrations (pg/ml) were normalized to cell number. ( C ) UPCI:SCC040. ( D ) Cal33. ( E-F ) Relative CXCR2 gene expression shown as ΔCT values in ( E ) Cal33 and ( F ) UPCI:SCC040 cells. Values a represent means ± SEM of N = 2. Student’s t test: p < 0.05 (*), p < 0.01 (**), ns: not significant

Article Snippet: Cytokines IL-1α, IL-1β, IL-8 and CXCL1 were measured using DuoSet ELISA kits (R&D Systems, IL-1α Cat#DY200, IL-1β Cat#DY201-05, IL8 Cat#30021151, CXCL1 Cat#DY 275) according to manufacturer instructions.

Techniques: Irradiation, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Correlations between LL-37 and citLL-37-mediated increase in the mRNA abundance of COX-2 and chemokines. HBEC-3KT cells were stimulated with either (⬤) LL-37 or (🞅) citLL-37 (0.50 µM each). mRNA abundance of COX-2, IL-8, GROα and MIP-3α were examined in cell lysates using qRT-PCR after 4 h. Relative fold changes were calculated compared to unstimulated cells normalized to 1, using the ΔΔCt method after normalization with 18s RNA expression. Pearson’s correlation analysis was performed to determine the correlation between COX-2 and ( a ) IL-8, ( b ) GROα and ( c ) MIP-3α mRNA abundance (fold changes compared to unstimulated cells)

Article Snippet: The abundance of IL-8 (Cat# DY208), GROα (Cat# DY275) and MIP-3α (Cat# DY360) were examined in TC supernatants using ELISA kits obtained from R&D Systems, according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, RNA Expression

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Inhibition of COX-2 suppresses LL-37-mediated chemokine production. HBEC-3KT cells were pre-treated with COX-2 inhibitor Rofecoxib (20 nM) for 1 h, followed by stimulation with either LL-37, citLL-37 or sLL-37 (0.50 µM each) for 24 h (n = 5). TC supernatants were examined for the abundance of ( a ) IL-8, ( b ) GROα and ( c ) MIP-3α by ELISA. Chemokine levels shown in pg/mL after background subtraction of levels in unstimulated cells. Each dot represents an independent experiment (n = 5), the bars show IQR with the median line and the whiskers represent the min-max range. Statistical significance was measured using Two-Way ANOVA, and # represents statistical significance compared to unstimulated cells ( ** p < 0.005, *** p < 0.0005, #### or **** p < 0.0001)

Article Snippet: The abundance of IL-8 (Cat# DY208), GROα (Cat# DY275) and MIP-3α (Cat# DY360) were examined in TC supernatants using ELISA kits obtained from R&D Systems, according to the manufacturer’s instructions.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Inhibition of PGE2 receptors (EP1-4) suppresses LL-37-mediated enhancement of chemokines. HBEC-3KT cells were pre-treated with specific inhibitors for PGE2 receptors, EP1 (SC-19220; 20 nM), EP2 (PF-044EP2; 25 nM), EP3 (L-798,106; 20 nM), or EP4 (MF498; 10 nM), 1 h prior to stimulation with either LL-37or sLL-37 (0.50 μM). Tissue culture (TC) supernatants were collected after 24 h and the abundance of ( a ) IL-8, ( b ) GROα and ( c ) MIP-3α were measured by ELISA. Each dot represents an independent experiment (n=4), the bars show IQR with the median line and the whiskers represent the min-max range. Results shown are after subtracting baseline values obtained from unstimulated cells in each independent experiment. Statistical significance was determined using Two-Way ANOVA (** p < 0.001 and **** p < 0.0001)

Article Snippet: The abundance of IL-8 (Cat# DY208), GROα (Cat# DY275) and MIP-3α (Cat# DY360) were examined in TC supernatants using ELISA kits obtained from R&D Systems, according to the manufacturer’s instructions.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Proposed mechanism of LL-37-COX-2 axis for chemokine production and neutrophil migration in the lungs. LL-37 engages the P2X 7 receptor in upregulating COX-2 expression which facilitates increase in the abundance of prostaglandin PGE2. Release of PGE2 may act in an autocrine manner through PGE2 receptors (EP1 − 4), resulting in enhanced production of chemokines including IL-8 and GROα, which facilitates neutrophil migration contributing to airway inflammation. However, citrullination of LL-37 dampens this pathway, potentially acting as a regulatory switch that limits the pro-inflammatory functions of LL-37 in the lungs (Figure created in BioRender.com)

Article Snippet: The abundance of IL-8 (Cat# DY208), GROα (Cat# DY275) and MIP-3α (Cat# DY360) were examined in TC supernatants using ELISA kits obtained from R&D Systems, according to the manufacturer’s instructions.

Techniques: Migration, Expressing