Journal: Acta Otorhinolaryngologica Italica

Article Title: Dose-dependent IL-29 activation of TLR4 signalling drives eosinophil infiltration in chronic rhinosinusitis with nasal polyps

doi: 10.14639/0392-100X-A1222

Figure Lengend Snippet: Effect of IL-29 on eosinophil inflammatory cytokine secretion. (A) ELISA analysis of IL-5 secretion in eosinophil supernatants; (B) ELISA analysis of IL-13 secretion in eosinophil supernatants. Eosinophils were stimulated with different concentrations of IL-29 (10, 25, 50, 100 ng/mL) for 24 hours. Data were obtained from 3 independent experiments and are presented as mean ± SD. * indicates p < 0.05.

Article Snippet: A total of 1×10 5 eosinophils were seeded in the upper chamber, while the lower chamber was filled with RPMI-1640 medium containing 10 ng/mL IL-5 (R&D Systems, 205-IL-010) as a chemoattractant.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Acta Otorhinolaryngologica Italica

Article Title: Dose-dependent IL-29 activation of TLR4 signalling drives eosinophil infiltration in chronic rhinosinusitis with nasal polyps

doi: 10.14639/0392-100X-A1222

Figure Lengend Snippet: The interventional effect of TAK-242 Blocking TLR4 on IL-29 action. (A) Transwell assay showing changes in eosinophil migration after TLR4 blockade with TAK-242; (B) ELISA of IL-5 secretion after TAK-242 intervention; (C) ELISA of IL-13 secretion after TAK-242 intervention; (D) Western blot analysis of TLR4, p-NF-κB, and p-MAPK protein expression after TAK-242 treatment. Data are presented as mean ± SD. * indicates p < 0.05.

Article Snippet: A total of 1×10 5 eosinophils were seeded in the upper chamber, while the lower chamber was filled with RPMI-1640 medium containing 10 ng/mL IL-5 (R&D Systems, 205-IL-010) as a chemoattractant.

Techniques: Blocking Assay, Transwell Assay, Migration, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Bromodomain and extra-terminal protein inhibitors modulate natural killer cell function and differentiation

doi: 10.1038/s41598-025-33437-1

Figure Lengend Snippet: BETi impact function of primary human NK cells. ( A ) Human primary NK cells isolated from healthy donors ( n = 4) unstimulated or stimulated with IL-12 + IL-15 for 24 h with prior (24 h) treatment with BETi (AZD5153 or CPI-203) compared to no drug treatment control. Bar graph of levels of IFNγ secreted into the cell culture supernatant measured by ELISA reported in pg/ml. ** P ≤ 0.01, * P ≤ 0.05, Ratio paired t-test. ( B ) Percentage of live CD14 − CD19 − cells and of total NK cells (live CD14 − CD19 − CD56 + CD16 + ) within the lymphocyte gate (Supplementary Fig. 2) in PBMCs from healthy donors ( n = 6) incubated in medium only (open circles) or with CPI-203 (black circles) or with AZD5153 (blue circles), in the presence or absence of IL-12 plus IL-18, for 48 h, and then co-cultured with CD4.221 cells for a further 5 h and analyzed. ( C ) Representative histogram overlays illustrating expression of granzyme B in NK cells in the indicated conditions, and bar graphs showing the median fluorescence intensity (MFI) of granzyme B staining in NK cells after culture of PBMCs from healthy donors ( n = 6) for 48 h in medium with or without IL-12 + IL-18 in the absence (open circles) or presence of BET inhibitors CPI-203 (black circles) or AZD5153 (blue circles), followed by co-culture for 5 h with CD4.221 cells in the presence of Brefeldin A. ( D – F ) Percentages of CD107a + ( D ), of IFN-γ + ( E ) and of TNF-α + cells ( F ) in total NK cells within PBMCs from healthy donors ( n = 6) incubated in medium only (open circles) or with CPI-203 (black circles) or with AZD5153 (blue circles), in the presence or absence of IL-12 plus IL-18, for 48 h, and then co-cultured with CD4.221 cells in the presence of Brefeldin A for a further 5 h and analyzed. In ( B – F ), each symbol represents data from a single donor; error bars indicate mean ± sem of n = 6 samples. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.00001; one way-ANOVA with Tukey’s post-test.

Article Snippet: Cells were stimulated with 10ng/ml recombinant IL-12 (130-129-720, Miltenyi) plus 100ng/ml recombinant IL-18 (B001-5, R&D Systems) and with 100nM CPI-203 (15479, Cayman Chemical) or 100nM AZD 5153 (15479, Cayman Chemical), or cultured in medium only for 48 h at 37 °C.

Techniques: Isolation, Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Incubation, Expressing, Fluorescence, Staining, Co-Culture Assay