anti il 36β  (R&D Systems)

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: IL-36α inhibits melanoma by inducing pro-inflammatory polarization of macrophages

doi: 10.1007/s00262-023-03477-5

Figure Lengend Snippet: IL-36α may have an anti-tumor function in both human and mice. a Relationship between high and low expression of IL-36A, IL-36B and IL-36G and patient survival in patients with thymoma, kidney renal clear cell carcinoma and melanoma. Data were from the Kaplan–Meier plotter database and the cBioPortal database. b Schematic protocol of the mice tumor model. Mice were challenged with 5 × 105 B16-vec or B16-IL-36α s.c. On day 14, melanoma tissue was removed and analyzed. c Mice weights were measured every 2 days from the fourth day after tumor implantation. (n = 5). Data are shown as mean ± SEM. Five mice were in each group. *p < 0.05; **p < 0.01; ***p < 0.001, determined by Mann–Whitney test. B16-IL-36α, B16-IL-36β and B16-IL-36γ was compared with B16-vec separately. d, f On day 14 after implantation, the mice tumors were removed and weighed. (n = 5). Data are shown as mean ± SEM. Five mice were in each group. *p < 0.05; **p < 0.01; ***p < 0.001, determined by one-way ANOVA with Dunn’s test. B16-IL-36α, B16-IL-36β and B16-IL-36γ were compared with B16-vec separately. e Tumor volume in mice was measured daily from day 8 after tumor implantation. (n = 5). Data are shown as mean ± SEM. Five mice were in each group. *p < 0.05; **p < 0.01; ***p < 0.001, determined by Mann–Whitney test. B16-IL-36α, B16-IL-36β and B16-IL-36γ were compared with B16-vec separately. g The survival of mice was monitored. Ten mice were in each group. The p value was based on a log-rank test. B16-IL-36α, B16-IL-36β and B16-IL-36γ were compared with B16-vec separately

Article Snippet: Protein expression levels of IL-36α, IL-36β, and IL-36γ were determined by western blotting using anti-IL-36α (AF2297-SP, R&D Systems), anti-IL-36β (PA5-81415, Invitrogen) and anti-IL-36γ (A10165, ABclonal) antibodies.

Techniques: Expressing, Tumor Implantation, MANN-WHITNEY

Journal: International journal of molecular sciences

Article Title: MES SV40 Cells Are Sensitive to Lipopolysaccharide, Peptidoglycan, and Poly I:C Expressing IL-36 Cytokines.

doi: 10.3390/ijms231911922

Figure Lengend Snippet: Figure 2. LPS stimulation triggers a differential expression of IL-36β in MCs. MES SV40 cells (1.5 × 106) were stimulated in 2 mL of medium containing different amounts of LPS for 2 h, and the expression of (A) IL-36α; (B) IL-36β; (C) IL-36γ mRNA was measured. The expression was normalized against Actb, and the fold change was calculated in comparison with the respective non-stimulated control (NTC). (D) Mesangial cells were stimulated for 6 h, and total protein was extracted for Western blot performance. The intensity of the bands was quantified, normalized against β-actin, and the fold change was calculated. E) Representative image of IL-36β production (stained green) in MES SV40 cells stimulated with 100 ng of LPS. The results were obtained from at least 3 independent experiments and are listed as mean ± SD. Data were analyzed with one-way ANOVA followed by Dunnett´s analysis. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the blocking solution was removed, 3 washes were performed with PBS, and the primary antibodies diluted 1:100 mouse anti-IL-36α/IL-1F6 (R&D SYSTEMS, Minneapolis, USA Catalog AF2297) or mouse anti IL-36β/IL-F8 (R&D SYSTEMS, Minneapolis, USA Catalog AF2298) were added.

Techniques: Quantitative Proteomics, Expressing, Comparison, Control, Western Blot, Staining

Journal: International journal of molecular sciences

Article Title: MES SV40 Cells Are Sensitive to Lipopolysaccharide, Peptidoglycan, and Poly I:C Expressing IL-36 Cytokines.

doi: 10.3390/ijms231911922

Figure Lengend Snippet: Figure 3. The stimulation of MCs with PGN generates the production of IL-36α. MES SV40 cells (1.5 × 106) were stimulated in 2 mL of medium containing different amounts of PGN for 6 h, and (A) the IL-36α; (B) IL-36β; (C) IL-36γ mRNA expressions were measured. The expression was normalized against Actb, and the fold change was calculated in comparison with the respective non-stimulated control (NTC). (D) Mesangial cells were stimulated under similar conditions, and total proteins were extracted for WB. The intensity of bands was normalized against β-actin, and the fold change was calculated. E) Representative IF image of IL-36α (green stained) in MCs stimulated with 1 µg of PGN for 6 h. Results were obtained from at least 3 independent experiments and are presented as mean ± SD. Data were analyzed with one-way ANOVA followed by Dunnett´s analysis. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the blocking solution was removed, 3 washes were performed with PBS, and the primary antibodies diluted 1:100 mouse anti-IL-36α/IL-1F6 (R&D SYSTEMS, Minneapolis, USA Catalog AF2297) or mouse anti IL-36β/IL-F8 (R&D SYSTEMS, Minneapolis, USA Catalog AF2298) were added.

Techniques: Expressing, Comparison, Control, Staining

Journal: International journal of molecular sciences

Article Title: MES SV40 Cells Are Sensitive to Lipopolysaccharide, Peptidoglycan, and Poly I:C Expressing IL-36 Cytokines.

doi: 10.3390/ijms231911922

Figure Lengend Snippet: Figure 4. Poly I:C promotes IL-36α expression in MCs. MES SV40 cells (1.5 × 106) were stimulated in 2 mL of medium containing different amounts of poly I:C for 2 h, and the expressions of (A) IL-36α; (B) IL-36β; (C) IL-36γ were measured. The expression was normalized against Actb, and the fold change was calculated against the non-stimulated control (NTC). (D) For the WB analysis, MCs were stimulated for 6 h under similar conditions, and total proteins were extracted. The intensity of bands was normalized against β-actin, and the fold change was calculated. (E) Representative IF image of IL-36α (green stained) in MCs stimulated with 0.1 ng of poly I:C for 6 h. The results were obtained from at least 3 independent experiments and are presented as mean ± SD. Data were analyzed with one-way ANOVA followed by Dunnett´s analysis. * p < 0.05, ** p < 0.01.

Article Snippet: Subsequently, the blocking solution was removed, 3 washes were performed with PBS, and the primary antibodies diluted 1:100 mouse anti-IL-36α/IL-1F6 (R&D SYSTEMS, Minneapolis, USA Catalog AF2297) or mouse anti IL-36β/IL-F8 (R&D SYSTEMS, Minneapolis, USA Catalog AF2298) were added.

Techniques: Expressing, Control, Staining