mouse interleukin 6 il 6 elisa kit  (Cusabio)

Journal: Molecular Medicine Reports

Article Title: Naringin ameliorates intestinal injury in ulcerative colitis model mice by modulating the JAK2/STAT3 signaling pathway

doi: 10.3892/mmr.2026.13805

Figure Lengend Snippet: Naringin inhibits JAK2/STAT3 signaling and restores tight junction proteins in dextran sulfate sodium-induced colitis. (A) Representative western blotting images of p-JAK2, JAK2, p-STAT3, STAT3, IL-6, occludin, ZO-1 and β-actin expression in colon tissues. Densitometric semi-quantification of the relative protein expression levels of (B) p-JAK2/β-actin, (C) JAK2/β-actin, (D) p-JAK2/JAK2, (E) p-STAT3/β-actin, (F) STAT3/β-actin, (G) p-STAT3/STAT3, (H) IL-6/β-actin, (I) occludin/β-actin and (J) ZO-1/β-actin. Data are presented as mean ± SEM. n=3. *P<0.05 and **P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. normal group. One-way ANOVA followed by Tukey's post-hoc test was applied. N, normal group; C, control group; Nar, naringin group; Mes, mesalazine group; p-, phosphorylated; JAK2, Janus kinase 2; STAT3, signal transducer and activator of transcription 3; IL-6, ZO-1, zona occludens-1.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 2 h at room temperature, then incubated at 4°C overnight with primary antibodies against IL-6 (1:1,000; cat. no. Bs-0782R; BIOSS), phosphorylated (p-)JAK2 (1:1,000; cat. no. bs-2485R; BIOSS), JAK2 (1:1,000; cat. no. bs-0908R; BIOSS), p-STAT3 (1:1,000; cat. no. bs-1658R; BIOSS), STAT3 (1:1,000; cat. no. bs-55208R; BIOSS), occludin (1:1,000; cat. no. A2601; ABclonal Biotech Co., Ltd.), zona occludens-1 (ZO-1; 1:1,000; cat. no. A0659; ABclonal Biotech Co., Ltd.) and β-actin (1:1,000; cat. no. ab8227; Abcam).

Techniques: Western Blot, Expressing, Control

Journal: Molecular Medicine Reports

Article Title: Naringin ameliorates intestinal injury in ulcerative colitis model mice by modulating the JAK2/STAT3 signaling pathway

doi: 10.3892/mmr.2026.13805

Figure Lengend Snippet: Naringin restores barrier function in IL-6-induced Caco-2 cells. (A) Transepithelial electrical resistance values over time. (B) Fluorescein isothiocyanate-dextran 4 kDa permeability assay. n=6. Data are presented as mean ± SEM. **P<0.01 vs. IL-6 group; ## P<0.01 vs. normal group. One-way ANOVA followed by Tukey's post-hoc test was applied. Nar, naringin.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 2 h at room temperature, then incubated at 4°C overnight with primary antibodies against IL-6 (1:1,000; cat. no. Bs-0782R; BIOSS), phosphorylated (p-)JAK2 (1:1,000; cat. no. bs-2485R; BIOSS), JAK2 (1:1,000; cat. no. bs-0908R; BIOSS), p-STAT3 (1:1,000; cat. no. bs-1658R; BIOSS), STAT3 (1:1,000; cat. no. bs-55208R; BIOSS), occludin (1:1,000; cat. no. A2601; ABclonal Biotech Co., Ltd.), zona occludens-1 (ZO-1; 1:1,000; cat. no. A0659; ABclonal Biotech Co., Ltd.) and β-actin (1:1,000; cat. no. ab8227; Abcam).

Techniques: Permeability

Journal: Molecular Medicine Reports

Article Title: Naringin ameliorates intestinal injury in ulcerative colitis model mice by modulating the JAK2/STAT3 signaling pathway

doi: 10.3892/mmr.2026.13805

Figure Lengend Snippet: Naringin reverses IL-6-induced ZO-1 and occludin impairment in a STAT3-dependent manner. (A) Immunofluorescence staining for ZO-1 (blue) and occludin (green); nuclei were stained with DAPI (cyan). Merged shows the merged images of DAPI and respective protein staining. Quantitative analysis of relative (B) ZO-1 and (C) occludin protein expression. (n=3). Data are presented as mean ± SEM. *P<0.05 and **P<0.01 vs. IL-6 group; ## P<0.01 vs. normal group. One-way ANOVA followed by Tukey's post-hoc test was applied. ZO-1, zona occludens-1; N, normal group; Nar, naringin; STAT3, signal transducer and activator of transcription 3; si, small interfering RNA; NC, negative control.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 2 h at room temperature, then incubated at 4°C overnight with primary antibodies against IL-6 (1:1,000; cat. no. Bs-0782R; BIOSS), phosphorylated (p-)JAK2 (1:1,000; cat. no. bs-2485R; BIOSS), JAK2 (1:1,000; cat. no. bs-0908R; BIOSS), p-STAT3 (1:1,000; cat. no. bs-1658R; BIOSS), STAT3 (1:1,000; cat. no. bs-55208R; BIOSS), occludin (1:1,000; cat. no. A2601; ABclonal Biotech Co., Ltd.), zona occludens-1 (ZO-1; 1:1,000; cat. no. A0659; ABclonal Biotech Co., Ltd.) and β-actin (1:1,000; cat. no. ab8227; Abcam).

Techniques: Immunofluorescence, Staining, Expressing, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: Naringin ameliorates intestinal injury in ulcerative colitis model mice by modulating the JAK2/STAT3 signaling pathway

doi: 10.3892/mmr.2026.13805

Figure Lengend Snippet: Naringin suppresses JAK2/STAT3 activation in IL-6-stimulated Caco-2 cells with STAT3 silencing. (A) Western blot analysis of p-JAK2, JAK2, p-STAT3, STAT3, occludin and ZO-1 in Caco-2 cells under indicated treatments. (B) Validation of STAT3 knockdown efficiency: Relative STAT3 expression in cells transfected with siSTAT3 vs. siNC. ## P<0.01 vs. siNC. Densitometric semi-quantification of relative protein expression levels of (C) p-JAK2/β-actin, (D) JAK2/β-actin, (E) p-JAK2/JAK2, (F) p-STAT3/β-actin, (G) STAT3/β-actin, (H) p-STAT3/ STAT3, (I) occludin/β-actin, (J) ZO-1/β-actin. Data are presented as mean ± SEM. n=3. *P<0.05 and **P<0.01 vs. IL-6 group; ## P<0.01 vs. normal group; Δ P<0.05 and ΔΔ P<0.01 IL-6 + Nar group vs. IL-6 + Nar + siSTAT3 group. One-way ANOVA followed by Tukey's post-hoc test was applied. N, normal group; Nar, naringin group; Mes, mesalazine group; p-, phosphorylated; JAK2, Janus kinase 2; STAT3, signal transducer and activator of transcription 3; ZO-1, zona occludens-1; si, small interfering RNA; NC, negative control.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 2 h at room temperature, then incubated at 4°C overnight with primary antibodies against IL-6 (1:1,000; cat. no. Bs-0782R; BIOSS), phosphorylated (p-)JAK2 (1:1,000; cat. no. bs-2485R; BIOSS), JAK2 (1:1,000; cat. no. bs-0908R; BIOSS), p-STAT3 (1:1,000; cat. no. bs-1658R; BIOSS), STAT3 (1:1,000; cat. no. bs-55208R; BIOSS), occludin (1:1,000; cat. no. A2601; ABclonal Biotech Co., Ltd.), zona occludens-1 (ZO-1; 1:1,000; cat. no. A0659; ABclonal Biotech Co., Ltd.) and β-actin (1:1,000; cat. no. ab8227; Abcam).

Techniques: Activation Assay, Western Blot, Biomarker Discovery, Knockdown, Expressing, Transfection, Small Interfering RNA, Negative Control

Journal: Bioactive Materials

Article Title: Bioengineered extracellular vesicles escape lysosomal degradation and deliver Tet-PKM2 for macrophage immunometabolic reprogramming and periodontitis treatment

doi: 10.1016/j.bioactmat.2026.01.002

Figure Lengend Snippet: Immunomodulatory effects of the bioengineered LEVs Tet−PKM2 @TA in terms of their ability to modulate macrophage polarization in vitro . The macrophages were treated with 100 ng/mL LPS for 24 h and then treated with PBS (Control), 100 μg/mL LEVs PKM2 , LEVs Tet−PKM2 , or LEVs Tet−PKM2 @TA for another 24 h. ( A ) The relative mRNA expression levels of M1 polarization-related genes ( IL-6 and IL-1β ) and M2 polarization-related genes ( IL-4 and Arg-1 ) in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups (qRT‒PCR) ( n = 3). ( B ) Concentrations of M1-related cytokines (IL-6 and TNF-α) and M2-related cytokines (IL-4 and IL-10) in the supernatants of the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups (ELISA) ( n = 3). ( C ) Representative immunofluorescence images and quantification of the expression levels of M1-related proteins (iNOS and CCR7) and M2-related proteins (CD163, CD206, and Arg-1) in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 3). The data are expressed as the mean ± SEM. Statistical analysis was performed with one-way ANOVA ( A , B , and C ). ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 indicate significant differences between the indicated columns.

Article Snippet: Kits were sourced as follows: TNF-α, IL-4, and IL-10 from Fankew (Shanghai Kexing Trading Co., Ltd., China) and IL-6 from Proteintech.

Techniques: In Vitro, Control, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: Exosomal HMGB1 Orchestrates NSCLC Progression and Immunosuppressive Macrophage Polarisation Through the TLR4 / NF ‐ κB / IL ‐6/ STAT3 Signalling Cascade

doi: 10.1111/jcmm.71050

Figure Lengend Snippet: Exosomal HMGB1 activates JAK/STAT3 signalling to promote NSCLC progression. (A) Protein–protein interaction (PPI) network analysis of HMGB1 using the STRING database. (B) Western blot analysis of NF‐κB in A549 and PC9 cells treated with PBS, recombinant HMGB1 (100 ng), exosomes from vector cells or exosomes from HMGB1 OE cells (cell‐to‐exosome ratio = 1:10). (C) ELISA quantification of IL‐6 in the supernatant of A549 and PC9 cells under the same treatment conditions as in (B). (D) Immunofluorescence staining of p‐STAT3 of A549 and PC9 cells under the same treatments, including an additional group co‐treated with exosomes from HMGB1 OE cells and NF‐κB inhibitor (50 μM). (E) Cell proliferation of A549 and PC9 cells treated with HMGB1 OE‐derived exosomes alone or in combination with NF‐κB inhibitor (50 μM) or STAT3 inhibitor (20 μM). (F) Cell migration under the same treatment conditions as in (E). (G) Colony formation assays of A549 and PC9 cells under the same treatment conditions as in (E).

Article Snippet: IL‐6 neutralising antibodies were obtained from Sino Biological (China).

Techniques: Western Blot, Recombinant, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Derivative Assay, Migration

Journal: Journal of Cellular and Molecular Medicine

Article Title: Exosomal HMGB1 Orchestrates NSCLC Progression and Immunosuppressive Macrophage Polarisation Through the TLR4 / NF ‐ κB / IL ‐6/ STAT3 Signalling Cascade

doi: 10.1111/jcmm.71050

Figure Lengend Snippet: Targeting HMGB1 signalling improves therapeutic outcomes in NSCLC. (A) Correlation analysis between immune infiltration scores and HMGB1 expression in 491 LUAD and 500 LUSC patients from the TCGA database. (B) Correlation between HMGB1 expression and the distribution of various immune cell subsets in LUAD and LUSC patients. (C, D) THP‐1–derived M0 macrophages were treated with PBS, HMGB1 (10 or 100 ng) or exosomes derived from vector or HMGB1 OE cells (cell‐to‐exosome ratio = 1:10). M1 macrophage markers (CD86, CD80, iNOS) and M2 markers (CD206, IL‐10, Arg1) were quantified by PCR. (E) Lewis tumour‐bearing mice were treated with PBS, HMGB1 OE‐derived exosomes (1 × 10 10 exosomes per mouse, twice per week), anti‐PD‐1 antibody (RMP1‐14, 200 μg per mouse, twice per week) or combination therapy ( n = 5 per group). Tumour volumes and apoptosis levels in tumour tissues (day 25) were assessed. (F) PC9 cells were treated with PBS or exosomes from HMGB1 OE cells (cell‐to‐exosome ratio = 1:10), followed by Osimertinib (50 nM, 48 h), and apoptosis was measured. (G) A549 and PC9 cells were similarly treated with PBS or HMGB1 OE‐derived exosomes, followed by Cisplatin (5 μM, 48 h), and apoptosis was analysed. (H) A549 and PC9 cells were similarly treated with paclitaxel (10 μM, 48 h) under the same conditions, and cell apoptosis was determined. (I) A549‐bearing mice were treated with HMGB1 OE‐derived exosomes (1 × 10 10 exosomes per mouse), followed by PBS, paclitaxel (PTX, 10 mg/kg, twice per week), STAT3 inhibitor (5 mg/kg, twice per week) or combination therapy. (J) Schematic diagram illustrating the proposed mechanism: HMGB1 upregulates TLR4, thereby activating the NF‐κB–IL‐6 axis and stimulating JAK2/STAT3 signalling to promote tumour progression. Concurrently, HMGB1 facilitates M2 macrophage polarisation.

Article Snippet: IL‐6 neutralising antibodies were obtained from Sino Biological (China).

Techniques: Expressing, Derivative Assay, Plasmid Preparation

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: rhIL-37 delays preterm delivery by reducing LPS-induced inflammatory effects and NLRP3 inflammasome activation in mice. ( A ) Orbital blood was collected from the control mice ( n = 6), LPS-induced preterm mice (LPS; n = 6), and rhIL-37 + LPS-co-treated preterm mice (rhIL-37 + LPS; n = 6).Serum levels of IL-1β, IL-6 and TNF-α were detected via ELISA. ( B ) Foetal membrane and placental tissues were collected from mice ( n = 6 per group) immediately before labour,, and the mRNA expression levels of NLRP3, Caspase-1 and ASC were detected via qRT-PCR. ( C ) Foetal membrane and placental tissues were collected from the mice in each group ( n = 3 per group), and the protein levels of NLRP3, Caspase-1, Pro-caspase-1 and ASC were analyzed by western blotting. ( D ) Western blot Results were normalised to β-actin. Bar graphs show statistical quantification of the western blot data. All data met the assumption of homogeneity of variance. Data are presented as mean ± s.e.m. Results are representative of at least three independent experiments. * p < 0.05; ** p < 0.01;*** p < 0.001;**** p < 0.0001 (one-way ANOVA).

Article Snippet: For mouse samples, IL-1β, IL-6, and TNF-α levels in orbital blood serum/plasma supernatants were assayed using a mouse ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Activation Assay, Control, Enzyme-linked Immunosorbent Assay, Membrane, Expressing, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: IL-37 knockdown enhances LPS-induced inflammatory and NLRP3 inflammasome activation in HTR-8/Svneo cells. ( A ) Cell supernatants were collected from control, si_NC- or si_IL37-treated cells following LPS exposure, and IL-37 expression level were detected by ELISA, qRT-PCR and Western blotting. ( B ) IL-1β, IL-6 and TNF-α levels in the cell supernatants were detected by ELISA. ( C ) The mRNA expression levels of NLRP3, Caspase-1 and ASC after cell treatment with LPS ( n = 5) were measured by qRT-PCR analysis. ( D ) NLRP3,pro-caspase-1, caspase-1 and ASC protein levels were detected by Western blotting, and the results were normalised to β-actin. All data exhibited equal variance.The data are shown as the mean ± s.e.m. The results are representative of at least three independent experiments. * p < 0.05; ** p < 0.01;*** p < 0.001;**** p < 0.0001 (One-way ANOVA).

Article Snippet: For mouse samples, IL-1β, IL-6, and TNF-α levels in orbital blood serum/plasma supernatants were assayed using a mouse ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Knockdown, Activation Assay, Control, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: IL-37 alleviates inflammatory effects and NLRP3 inflammasome activation in LPS-induced preterm birth

doi: 10.1038/s41598-025-34506-1

Figure Lengend Snippet: Nigericin reversed the anti-inflammatory effects and effects of rhIL-37 and its inhibition of NF-κB p65 activation in LPS-induced PTB.t ( A ) Cell supernatants were collected from HTR-8/Svneo cells co-treated with rhIL-37 and LPS, then either left untreated or treated with nigericin.The mRNA expression of NLRP3, caspase-1, and ASC was detected. ( B ) Cells were collected, and the protein expression of NLRP3, caspase-1, pro-caspase-1, and ASC was detected by western blotting. Results were normalised to β-actin and statistically analyzed.( C ) HTR-8/Svneo cell supernatants were collected after co-treatment with rhIL-37 and LPS (with or without nigericin), and the expression levels of IL-1β, IL-6, and TNF-α were detected by ELISA. ( D ) HTR-8/Svneo cells co-treated with rhIL-37 and LPS (with or without nigericin) were collected. The mRNA expression of p65 was detected by qRT-PCR, and protein expression of p65 and p-p65 was detected by western blotting. Densitometric quantification of p-p65 relative to p65 was performed. Statistical analysis of the western blot results was performed.All data exhibited equal variance.The data are shown as the means ± s.e.m. The results are representative of at least three independent experiments..

Article Snippet: For mouse samples, IL-1β, IL-6, and TNF-α levels in orbital blood serum/plasma supernatants were assayed using a mouse ELISA kit (LiankeBio, Hangzhou, China).

Techniques: Inhibition, Activation Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR