recombinant mouse il-33 109-266 protein (ril-33  (Enzo Biochem)

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 was upregulated and positively correlated with NET formation in renal transplant patients. ( A ) Paired analysis of serum IL-33 levels, ( B ) paired analysis of serum MPO-DNA levels, ( C ) paired analysis of serum citH3 levels in renal transplant patients preoperatively and postoperatively ( n = 20). ( D ) Correlation analysis for postoperative serum IL-33 level and serum MPO-DNA level (Spearman r = 0.6892, P = 0.0008). ( E ) Correlation analysis of postoperative serum IL-33 level and serum citH3 level (Spearman r = 0.6486, P = 0.002). * P < 0.05, ** P < 0.01

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques:

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 and NETs were elevated following renal I/R in mice. ( A - B ) Serum IL-33 levels ( A ) and renal tissue homogenate IL-33 levels ( B ) after renal I/R ( n = 6). ( C - D ) Representative blots ( C ) and statistical analysis ( D ) of IL-33 protein expression levels in renal tissues after I/R ( n = 6). ( E - F ) Representative images ( E ) and statistical analysis ( F ) of IL-33 immunohistochemistry in renal tissues after I/R ( n = 6). Scale bar was 50 μm. ( G - H ) IL-33 expression and distribution among the groups (blue staining indicate DAPI for nuclei), IL-33 (red) and CD31 (green) ( n = 6). Scale bar was 50 μm. (I-J) Serum MPO-DNA (I) and citH3 levels ( J ) following renal I/R ( n = 6). ( K - L ) Representative blots ( K ) and statistical analysis ( L ) of citH3 protein expression levels in renal tissues after I/R ( n = 6). ( M - N ) NET formation in in the indicated groups (blue indicate DAPI staining), MPO (red) and citH3 (green) ( n = 6). Scale bar was 20 μm. ns no significance, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 promoted NET generation during renal IRI. ( A ) A diagram showing WT mice receiving renal IRI or sham surgery following intraperitoneal injection of rIL-33 (10 µg per mouse) or vehicle (PBS). ( B - C ) Serum MPO-DNA ( B ) and citH3 levels ( C ) of mice in each group ( n = 6). ( D - E ) Representative blots ( D ) and statistical analyses ( E ) of citH3 protein expression levels in renal tissues in the indicated groups ( n = 6). ( F - G ) NET formation in the indicated groups through DAPI staining (blue), MPO (red) and citH3 (green) ( n = 6). Scale bar was 20 μm. ( H - I ) Serum Cr ( H ) and BUN levels ( I ) in the indicated groups ( n = 6). ( J - K ) Representative blots ( J ) and statistical analysis ( K ) of KIM-1 protein expression levels in renal tissues in the indicated groups ( n = 6). ( L - N ) Renal tissues from mice in the indicated groups stained with HE and KIM-1 ( L ), followed by score of tubular injury ( M ) and quantitative analysis of KIM-1 ( N ) ( n = 6). Scale bar was 50 μm. * P < 0.05, ** P < 0.01

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Injection, Expressing, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 exacerbated renal IRI by inducing NET formation. ( A ) A diagram showing the time node of intraperitoneal injection of GSK484 (4 mg/kg) and rIL-33 (10 µg per mouse) in WT mice before renal I/R. (B-C) Serum MPO-DNA ( B ) and citH3 levels ( C ) in the indicated groups ( n = 6). ( D - E ) Representative blots ( D ) and statistical analysis ( E ) of citH3 expression levels in renal tissues of the indicated groups ( n = 6). ( F - G ) NET formation in the indicated groups as determined using the DAPI (blue), MPO (red) and citH3 (green) staining ( n = 6). Scale bar was 20 μm. ( H - I ) Serum Cr ( H ) and BUN levels ( I ) in the indicated groups ( n = 6). ( J - K ) Representative blots ( J ) and statistical analysis ( K ) of KIM-1 protein expression levels in renal tissues from the indicated groups ( n = 6). ( L - N ) Renal tissues of mice in the indicated groups stained with HE and KIM-1 ( L ), followed by score of tubular injury ( M ) and quantitative analysis of KIM-1 ( N ) ( n = 6). Scale bar was 50 μm. ** P < 0.01, *** P < 0.001

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Injection, Expressing, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: Blocking IL-33 improved renal IRI by reducing NET formation. ( A ) The diagram of WT mice receiving renal IRI or sham surgery after intraperitoneal injection of anti-IL-33 monoclonal antibody (Anti-IL-33) (10 µg per ounce) or vehicle (IgG2b Isotype Control). ( B - C ) Serum MPO-DNA ( B ) and citH3 levels ( C ) of mice in the indicated groups ( n = 6). (D-E) Representative blots ( D ) and statistical analysis ( E ) of citH3 protein expression levels in renal tissues in the indicated groups ( n = 6). ( F - G ) NET formation in the indicated groups following DAPI (blue), MPO (red) and citH3 (green) staining ( n = 6). Scale bar was 20 μm. ( H - I ) Serum Cr ( H ) and BUN levels ( I ) in the indicated group ( n = 6). ( J - K ) Representative blots ( J ) and statistical analysis ( K ) of KIM-1 protein expression levels in renal tissues in the indicated groups ( n = 6). ( L - N ) Renal tissues of mice in the indicated groups subjected to HE and KIM-1 staining ( L ), followed by score of tubular injury ( M ) and statistical analysis of KIM-1 ( N ) ( n = 6). Scale bar was 50 μm. ** P < 0.01, *** P < 0.001

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Blocking Assay, Injection, Control, Expressing, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 stimulated NET generation in vitro. ( A - B ) Neutrophils incubated with PBS, PMA (100nM) and various concentrations of rIL-33 (20, 60, 100 ng/mL) for 4 h. rIL-33 stimulation increased MPO-DNA ( A ) and citH3 levels ( B ) in the neutrophil culture medium in a dose-dependent manner relative to the PBS group ( n = 3). ( C - D ) Relative to the PBS group, rIL-33 activated neutrophils to increase citH3 protein in a dose-dependent manner ( n = 3). ( E ) Compared with the PBS group, rIL-33 increased the expression of ST2 mRNA on neutrophils ( n = 3). ( F ) Representative scanning electron microscopy graphs of neutrophils treated with PBS or rIL-33 (100ng/mL) for 4 h. Scale bar was 10 μm. ( G ) The NET formation as indicated by DAPI (blue), MPO (red) and citH3 (green) staining, was comparable between rIL-33 (100ng/mL) and PMA group. Scale bar was 50 μm. ns no significance, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: In Vitro, Incubation, Expressing, Electron Microscopy, Staining

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: IL-33 induced NET formation via ST2/PI3K/Akt and ST2/PAD4 signaling pathways. Mouse bone marrow-derived neutrophils were treated with PBS or rIL-33 (100ng/mL) for 4 h, followed by RNA sequencing ( n = 3). ( A - B ) Volcano plot ( A ) and heatmap ( B ) showing differential gene expression between PBS and rIL-33 groups. ( C ) GO enrichment analysis of DEGs. ( D ) KEGG pathway enrichment analysis of DEGs. Bone marrow-derived neutrophils from WT and ST2 KO mice were treated with PBS or rIL-33 (100ng/mL) for 4 h. The production of MPO-DNA ( E ) and citH3 ( F ) in the cell culture medium of ST2 KO neutrophils treated with IL-33 was markedly decreased relative to the IL-33-treated WT neutrophils ( n = 3). ( G ) Confocal microscopy was conducted to examine NET formation (co-localization of DAPI, MPO and citH3) in each group. ( H - I ) The expression of PI3K, p-PI3K, Akt, p-Akt and citH3 in WT and ST2 KO neutrophils stimulated by rIL-33 as determined by Western blot ( n = 3). ( J - K ) WT neutrophils were treated with 10µM LY294002 (PI3K inhibitor) or 10µM MK2206 (Akt inhibitor) for 24 h, and 100ng/mL rIL-33 was added on the 20th h to stimulate WT neutrophils for 4 h. The expression of PI3K, p-PI3K, Akt, p-Akt and citH3 in neutrophils was determined by Western blot ( n = 3). ( L ) Gene expression of Padi1, Padi2, Padi4 and Padi6 in neutrophils stimulated with rIL-33 ( n = 3). The gene expression data were expressed as log 2 (FPKM + 1). ( M - N ) PAD4 protein expression levels and quantitative analysis in WT and ST2 KO neutrophils treated with rIL-33 were quantified by Western blots ( n = 3). ( O - P ) WT neutrophils were pretreated with GSK484 (PAD4 inhibitor) for 30 min after treatment with 100ng/mL rIL-33 for 4 h. The protein expression of citH3 in neutrophils was determined by Western blot ( n = 3). ns no significance, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques: Protein-Protein interactions, Derivative Assay, RNA Sequencing, Gene Expression, Cell Culture, Confocal Microscopy, Expressing, Western Blot

Journal: Inflammation

Article Title: Interleukin-33 Promotes Neutrophil Extracellular Trap Formation To Aggravate Renal Ischemia-Reperfusion Injury Through ST2/PI3K/Akt and ST2/PAD4 Pathways

doi: 10.1007/s10753-025-02364-8

Figure Lengend Snippet: Schematic diagram of the mechanism by which IL-33 exacerbated renal IRI by enhancing NET formation during renal I/R

Article Snippet: Mice were intraperitoneally injected with recombinant IL-33 (rIL-33) (10ug per mouse, MedChemExpress, America) or PBS immediately following exposure to renal ischemia.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: IL-33/ST2 Axis Plays a Protective Effect in Streptococcus pyogenes Infection through Strengthening of the Innate Immunity

doi: 10.3390/ijms221910566

Figure Lengend Snippet: The bacterial loads, the numbers, and the cell viability of infiltrating cells in the air pouches of GAS-infected WT, IL - 33 -KO and ST2 -KO mice. Groups of four WT, IL - 33 -KO and ST2 -KO mice were inoculated via the intra-air pouch route with 2 × 10 8 S. pyogenes NZ131 cells per mouse. In one group, IL - 33 -KO mice were treated with rIL-33 (1 µg per mouse) via the intra-air pouch route at 30 min before GAS infection and 24 h post-GAS infection. In the other group, WT mice were treated with sST2 fusion protein (1 µg per mouse) via the intra-air pouch route at 30 min before GAS infection and 24 h post-GAS infection. At 24 h or 48 h post-infection, exudates of the air pouch were collected, and bacterial numbers ( a ), the numbers ( b ) as well as the cell viability ( c ) of infiltrating cells were counted, as described in Materials and Methods. The results are expressed as the mean ± standard deviation. The infiltrating cells in the air pouches of WT, IL - 33 -KO and ST2 -KO mice given PBS only, without GAS infection, were approximately (1.5 ± 0.3) × 10 4 cells/mL. The infiltrating cells in the air pouch of WT mice given sST2 were approximately (2.5 ± 0.4) × 10 4 cells/mL, while infiltrating cell numbers of B6 mice given rIL-33 were approximately (1.5 ± 0.5) × 10 6 cells/mL. Treatment groups were compared for significance using ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with the GAS-infected WT group.

Article Snippet: In some experiments, 0.1 mL of mouse rIL-33 (R&D Systems) (1 µg per mouse) and sST2 protein (R&D Systems) (1 µg per mouse) were injected into the air pouches of IL - 33 -KO mice and WT mice, respectively, at 30 min before GAS infection or 24 h post-infection.

Techniques: Infection, Standard Deviation