stat3  (MedChemExpress)

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: For drug resistance assays, mice were treated with paclitaxel (10 mg/kg, i.p., twice a week), with or without STAT3 inhibitor (STAT3‐IN‐3, 5 mg/kg, i.p., MedChemExpress, USA).

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: For drug resistance assays, mice were treated with paclitaxel (10 mg/kg, i.p., twice a week), with or without STAT3 inhibitor (STAT3‐IN‐3, 5 mg/kg, i.p., MedChemExpress, USA).

Techniques: Expressing, Derivative Assay, Plasmid Preparation

Journal: European Journal of Medical Research

Article Title: PEAK1 promotes prostate cancer progression and docetaxel resistance by mediating the polarization of tumor-associated macrophages

doi: 10.1186/s40001-025-03568-2

Figure Lengend Snippet: Functions of PEAK1 on the HIF-1α/STAT3/NF-κB pathway activation in PCa cells. A WB was conducted to test the protein levels of HIF-1α, PD-L1, p -STAT3, and p –p65 in PC3 and DU145 cells with PEAK1 overexpression. B , C WB was conducted to test the protein levels of HIF-1α, PD-L1, p -STAT3, and p –p65 in PC3 and DU145 cells with PEAK1 knocking down. D WB was conducted to test the protein levels of HIF-1α, PD-L1, p -STAT3, and p –p65 in PC3 cells with PEAK1 overexpression and co-culturing with THP1 cells. E , F WB was conducted to test the protein levels of HIF-1α, PD-L1, p -STAT3, and p –p65 in PC3 and DU145 cells with PEAK1 overexpression and LW6, Stattic or Bay 11–7082 treatment. N = 3. * stands for p < 0.05, ** stands for p < 0.01, *** stands for p < 0.001

Article Snippet: The PC3 and DU145 cells were treated with the HIF-1α inhibitor LW6 (10 μM, #HY-13671, MedChemExpress) [ ]; the STAT3 inhibitor Stattic (10 μM, #HY-13818, MedChemExpress) [ ]; the NF-κB inhibitor Bay 11–7082 (15 μM, #HY-13453, MedChemExpress) [ ]; and increasing doses of enzalutamide (5 μM, 10 μM, 20 μM, and 40 μM, #HY-70002, MedChemExpress) [ ] for 24 h.

Techniques: Activation Assay, Over Expression

Journal: European Journal of Medical Research

Article Title: PEAK1 promotes prostate cancer progression and docetaxel resistance by mediating the polarization of tumor-associated macrophages

doi: 10.1186/s40001-025-03568-2

Figure Lengend Snippet: Mechanistic diagram of PEAK1 in PCa. Summary diagram of the functions and mechanisms of PEAK1 in PCa. Upregulated PEAK1 promotes the HIF-1α/STAT3/NF-κB pathway activation, thus enhancing tumor proliferation, migration, EMT, and reducing apoptosis, docetaxel/enzalutamide sensitivity. PEAK1 enhances CCL2, IL-6 and PD-L1 expression from PCa cells, which mediate “M2” polarization of TAMs and inducing TGF- β expression from TAMs. TGF- β can act on PCa cells and induce PEAK1 upregulation

Article Snippet: The PC3 and DU145 cells were treated with the HIF-1α inhibitor LW6 (10 μM, #HY-13671, MedChemExpress) [ ]; the STAT3 inhibitor Stattic (10 μM, #HY-13818, MedChemExpress) [ ]; the NF-κB inhibitor Bay 11–7082 (15 μM, #HY-13453, MedChemExpress) [ ]; and increasing doses of enzalutamide (5 μM, 10 μM, 20 μM, and 40 μM, #HY-70002, MedChemExpress) [ ] for 24 h.

Techniques: Activation Assay, Migration, Expressing

Journal: Communications Biology

Article Title: L-cystine alleviates necrotizing enterocolitis by regulating ferroptosis and Th17 cell differentiation via the IL-6/STAT3 pathway

doi: 10.1038/s42003-025-09438-1

Figure Lengend Snippet: A The standard chemical structure of L-cystine. B Cell viability measured by CCK-8 assay. C IL-6 mRNA and protein levels analyzed by qRT-PCR and Western blot. D IL-6 concentration in cell supernatant quantified by ELISA. E ROS generation assessed via flow cytometry. F Intracellular Fe²⁺ levels. G 4-HNE expression detected by immunofluorescence. Scale bar = 25 μm. H , I MDA, GSH, and SOD levels. J Western blot analysis of STAT3 phosphorylation and NCOA4/FTH1 protein levels. n = 3 biologically independent samples. Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For therapeutic intervention, pups received daily oral L-cystine (200 mg/mL in saline solution; C6H12N2O4S2, 56-89-3, MCE, Monmouth Junction, NJ, USA) and intraperitoneal injections of the STAT3 inhibitor S3I-201 (0.5 mg/mL in DMSO and 20% cyclodextrin; 501919-59-1, MCE) twice weekly .

Techniques: CCK-8 Assay, Quantitative RT-PCR, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Immunofluorescence, Phospho-proteomics

Journal: Communications Biology

Article Title: L-cystine alleviates necrotizing enterocolitis by regulating ferroptosis and Th17 cell differentiation via the IL-6/STAT3 pathway

doi: 10.1038/s42003-025-09438-1

Figure Lengend Snippet: A Cell viability assessed by CCK-8. B Fe²⁺ levels. C 4-HNE expression detected by immunofluorescence. Scale bar = 25 μm. D ROS generation via flow cytometry. E MDA, GSH, and SOD levels. F STAT3 phosphorylation and NCOA4/FTH1 levels using Western blot. n = 3 biologically independent samples. Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For therapeutic intervention, pups received daily oral L-cystine (200 mg/mL in saline solution; C6H12N2O4S2, 56-89-3, MCE, Monmouth Junction, NJ, USA) and intraperitoneal injections of the STAT3 inhibitor S3I-201 (0.5 mg/mL in DMSO and 20% cyclodextrin; 501919-59-1, MCE) twice weekly .

Techniques: CCK-8 Assay, Expressing, Immunofluorescence, Flow Cytometry, Phospho-proteomics, Western Blot

Journal: Communications Biology

Article Title: L-cystine alleviates necrotizing enterocolitis by regulating ferroptosis and Th17 cell differentiation via the IL-6/STAT3 pathway

doi: 10.1038/s42003-025-09438-1

Figure Lengend Snippet: A Cell viability measured by CCK-8. B ROS generation via flow cytometry. C Fe²⁺ levels. D 4-HNE expression detected by immunofluorescence. Scale bar = 25 μm. E MDA, GSH, and SOD levels. F IL-6 concentration in supernatant by ELISA. G STAT3 phosphorylation and NCOA4/FTH1 levels using Western blot. n = 3 biologically independent samples. Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For therapeutic intervention, pups received daily oral L-cystine (200 mg/mL in saline solution; C6H12N2O4S2, 56-89-3, MCE, Monmouth Junction, NJ, USA) and intraperitoneal injections of the STAT3 inhibitor S3I-201 (0.5 mg/mL in DMSO and 20% cyclodextrin; 501919-59-1, MCE) twice weekly .

Techniques: CCK-8 Assay, Flow Cytometry, Expressing, Immunofluorescence, Concentration Assay, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Western Blot

Journal: Communications Biology

Article Title: L-cystine alleviates necrotizing enterocolitis by regulating ferroptosis and Th17 cell differentiation via the IL-6/STAT3 pathway

doi: 10.1038/s42003-025-09438-1

Figure Lengend Snippet: A Flow cytometry sorting of native CD4 + T cells from patient peripheral blood. B , C Native CD4 + T cells treated with NCM-460-conditioned medium (CM) and anti-IL-6 antibody (900 nM) for 48 h. Flow cytometry analysis of Treg/Th17 populations and ratios. D , E Native CD4 + T cells treated with gradient concentrations of recombinant IL-6 (rIL-6). Flow cytometry analysis of Treg/Th17 populations and ratios. F , G Native CD4 + T cells treated with NCM-460-CM, rIL-6 (20 ng/mL), and STAT3 inhibitor S3I-201 (50 µM) for 48 h. Flow cytometry analysis of Treg/Th17 populations and ratios. H Western blot analysis of STAT3 phosphorylation. n = 3 biologically independent samples. Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For therapeutic intervention, pups received daily oral L-cystine (200 mg/mL in saline solution; C6H12N2O4S2, 56-89-3, MCE, Monmouth Junction, NJ, USA) and intraperitoneal injections of the STAT3 inhibitor S3I-201 (0.5 mg/mL in DMSO and 20% cyclodextrin; 501919-59-1, MCE) twice weekly .

Techniques: Flow Cytometry, Recombinant, Western Blot, Phospho-proteomics

Journal: Communications Biology

Article Title: L-cystine alleviates necrotizing enterocolitis by regulating ferroptosis and Th17 cell differentiation via the IL-6/STAT3 pathway

doi: 10.1038/s42003-025-09438-1

Figure Lengend Snippet: A MDA, GSH, and SOD levels. B Flow cytometry analysis of ROS generation. C Fe²⁺ levels. D Mitochondrial morphology in colon tissues using TEM. Scale bar = 500 nm. E , F Immunofluorescence showing 4-HNE expression in colon tissues. Scale bar = 25 μm. G Western blot analysis of STAT3 phosphorylation and NCOA4/FTH1 levels. n = 3 animals (both sexes). Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For therapeutic intervention, pups received daily oral L-cystine (200 mg/mL in saline solution; C6H12N2O4S2, 56-89-3, MCE, Monmouth Junction, NJ, USA) and intraperitoneal injections of the STAT3 inhibitor S3I-201 (0.5 mg/mL in DMSO and 20% cyclodextrin; 501919-59-1, MCE) twice weekly .

Techniques: Flow Cytometry, Immunofluorescence, Expressing, Western Blot, Phospho-proteomics