il4  (R&D Systems)

Journal: bioRxiv

Article Title: Curcumin Alleviates Systemic Inflammation and Gut Dysbiosis Induced by Circadian Rhythm Disruption in a Rodent Model of Jet Lag

doi: 10.64898/2026.01.18.698756

Figure Lengend Snippet: Schematic in the left panel shows that animals were individually housed in cages equipped with running wheels, and wheel revolutions were monitored daily using automated software. Rats were fed either curcumin or almond butter (vehicle) diet balls. Schematic in the right panel shows the timeline and overview of experimental procedures. Following behavioral testing, blood samples were collected for ELISA-based cytokine and chemokine analysis. Small intestine segments were harvested for Peyer’s patch analysis using ImageJ. Fecal samples were collected for microbiome analysis.

Article Snippet: Quantikine® high-sensitivity colorimetric sandwich ELISA kits (R&D Systems) were used to quantify levels of tumor necrosis factor alpha (TNF-α; RTA00), interferon gamma (IFN-γ; RIF00), interleukin-6 (IL-6; R6000B), cytokine-induced neutrophil chemoattractant-1 (CXCL1/CINC-1; RCN100), and interleukin-4 (IL-4; R4000).

Techniques: Software, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Influence of the Cholinergic System on the Pathogenesis of Glioblastoma: Impact of the Neutrophil Granulocytes

doi: 10.3390/ijms27010321

Figure Lengend Snippet: Influence of the cholinergic system on IL-8 production by U251 tumor cells. ( A ) IL-8 levels in culture supernatants measured by ELISA after culture. Ten experiments were performed in the 2D model (** p < 0.005, mean ± SD). ( B ) Correlation of CHRM1 (upper panels) and CHRM3 (lower panels) and of ( C ) ChAT (upper panels) and AChE (lower panels) with IL-8 expression in LGG (n = 516) and GBM (n = 153) samples, estimated by TIMER2.0 ( http://timer.cistrome.org/ ). ( D – G ) PD-L1 expression in U251 tumor cells. ( D ) PD-L1 expression in U251 cells in 2D model (monolayer grown for 24 h with cholinergic system agonists ACh (10 −8 –10 −9 M) or Carb (10 −8 –10 −9 M) and ( F ) 3D model (spheroids grown for 9 days in 1.5% agar, 5 × 10 4 cells/mL, treated with ACh (10 −8 –10 −9 M) or Carb (10 −8 –10 −9 M). * p < 0.05. ( E ) Representative experiment on 2D cultures and ( G ) 3D cultures. ( H ) Correlation of CHRM1 (upper panels) and CHRM3 (lower panels), and of ( I ) ChAT (upper panels) and AChE (lower panels), with PD-L1 expression in LG (n = 516) and GBM (n = 153) samples, estimated by TIMER2.0.

Article Snippet: Human IL-8 ELISA kit was obtained from R&D Systems (Minneapolis, MN, USA), and VEGF ELISA kit was obtained from Biolegend (San Diego, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Influence of the Cholinergic System on the Pathogenesis of Glioblastoma: Impact of the Neutrophil Granulocytes

doi: 10.3390/ijms27010321

Figure Lengend Snippet: Relationship between VEGF expression and the cholinergic system in GBM. ( A ) Correlation of CHRM1 (upper panels) and CHRM3 (lower panels) with VEGF-A expression in LGG (n = 516) and GBM (n = 153) samples, estimated by TIMER2.0 ( http://timer.cistrome.org/ ). ( B ) Correlation of ChAT (upper panels) and AChE (lower panels) with VEGF-A expression in LGG and GBM samples, estimated by TIMER2.0. ( C – G ) VEGF production in U251 cells cultured as 3D spheroids (9 days, 1.5% agar, 5 × 10 4 cells/mL) and treated with ACh (10 −8 –10 −9 M), Carb (10 −8 –10 −9 M), Tio (30 nM, 15 min pretreatment followed by ACh or Carb 10 −8 M), or Riv (5 μM). Supernatants were collected and VEGF levels measured by ELISA. Nine independent experiments were performed (** p < 0.005, mean ± SD). ( H – L ) VEGF production in U251 cells cultured as 2D monolayers (24 h) with the same treatments described above. Ten independent experiments were performed (* p < 0.05, ** p < 0.005, *** p < 0.0005, **** p < 0.0001, mean ± SD). ( M ) ICAM-1 expression in 3D cultures measured by flow cytometry. A representative experiment is shown.

Article Snippet: Human IL-8 ELISA kit was obtained from R&D Systems (Minneapolis, MN, USA), and VEGF ELISA kit was obtained from Biolegend (San Diego, CA, USA).

Techniques: Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Influence of the Cholinergic System on the Pathogenesis of Glioblastoma: Impact of the Neutrophil Granulocytes

doi: 10.3390/ijms27010321

Figure Lengend Snippet: Functional evaluation of the cholinergic system in PMN-p, PMN-h and cocultures with the U251 cell line. ( A – C ) PD-L1 expression in PMN-h cultured (5 × 10 6 cells/mL) with or without U251 cells for 24 h in the presence of ACh (10 −8 M) or Carb (10 −8 M). PD-L1 levels were measured by flow cytometry using CD11b as PMN marker. ( D , E ) PD-L1 expression in PMN-p cultured for 24 h with ACh (10 −8 –10 −9 M). Seven independent experiments were performed (** p < 0.005). ( J , K ) PD-L1 expression in PMN-h cultured under the same conditions. ( E , K ) Representative experiment. ( F , G , L , M ) CD11b expression in ( F , G ) PMN-p and ( L , M ) PMN-h after 15 min stimulation with ACh (10 −8 –10 −9 M). CD11b levels were evaluated by flow cytometry. Seven independent experiments were performed (* p < 0.05). ( G , M ) Representative experiments. ( H , I , N , O ) IL-8 and VEGF production in ( H , I ) PMN-p ( N , O ) and PMN-h after 24 h stimulation with ACh (10 −8 –10 −9 M). Cytokine levels were measured in culture supernatants by ELISA. Seven independent experiments were performed (* p < 0.05).

Article Snippet: Human IL-8 ELISA kit was obtained from R&D Systems (Minneapolis, MN, USA), and VEGF ELISA kit was obtained from Biolegend (San Diego, CA, USA).

Techniques: Functional Assay, Expressing, Cell Culture, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay