p stat3 inhibitor stattic  (MedChemExpress)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Tumor suppressor SLC9A2 inhibits colorectal cancer metastasis and reverses immunotherapy resistance by suppressing angiogenesis

doi: 10.1186/s13046-025-03422-7

Figure Lengend Snippet: SLC9A2 inhibits the migration and invasion of CRC cells by suppressing the STAT3 signaling pathway. ( A ) Pathway enrichment analysis was conducted to compare the SLC9A2-overexpressing CRC cells with the control group. ( B ) Western blot analysis was performed to measure the levels of SLC9A2, STAT3 and p-STAT3 Y705 in HCT116 cells transfected with either siRNA-NC or siRNA-SLC9A2. ( C ) Western blot analysis was performed to measure the levels of SLC9A2, STAT3 and p-STAT3 Y705 in HCT116-Hm cells overexpressing either Vector or SLC9A2. ( D ) HCT116 cells were transfected with either siRNA-NC or siRNA-SLC9A2, followed by treatment with or without 5 µM Stattic for 24 h. After 48 h, Western blot analysis was performed to evaluate the levels of STAT3 and p-STAT3 Y705 . ( E-F ) HCT116 cells were transfected with either siRNA-NC or siRNA-SLC9A2, and the siRNA-SLC9A2 group was subsequently treated with 5 µM Stattic for 24 h. Following a 48-hour incubation, nuclear-cytoplasmic separation was performed, and Western blot analysis was conducted to evaluate the levels of STAT3 and p-STAT3 Y705 ( E ). Immunofluorescence was used to assess the nuclear localization of p- STAT3 Y705 ( F ). ( G ) HCT116-Hm cells were transfected with either vector or SLC9A2. After 24 h, immunofluorescence was used to examine the nuclear localization of p-STAT3 Y705 . ( H ) Migration and invasion assays were conducted on HCT116 cells transfected with either siRNA-NC or siRNA-SLC9A2, and the siRNA-SLC9A2 group was subsequently treated with 5 µM Stattic for 24 h. ( I ) Wound healing assay was conducted on HCT116 cells transfected with either siRNA-NC or siRNA-SLC9A2, and the siRNA-SLC9A2 group was subsequently treated with 5 µM Stattic for 24 h. ( J ) Schematic diagram illustrating SLC9A2 knockdown and overexpression in PDOs from primary CRC and liver metastases to evaluate STAT3 activation. ( K ) Western blot analysis of p-STAT3 Y705 levels in PDOs with SLC9A2 knockdown from primary CRC (left) and overexpression from liver metastases (right). ( L ) Western blot analysis of p-STAT3 Y705 levels in mouse liver metastases as shown in Fig. C. Data in bar graphs indicate mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. Multi-group analysis of variance ( B , H , I ), Student’s t test ( C , L )

Article Snippet: The p-STAT3 Y705 inhibitor Stattic (MCE, HY-13818) and the JAK1/2 inhibitor Ruxolitinib (MCE, HY-50856) were utilized in this study.

Techniques: Migration, Control, Western Blot, Transfection, Plasmid Preparation, Incubation, Immunofluorescence, Wound Healing Assay, Knockdown, Over Expression, Activation Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Tumor suppressor SLC9A2 inhibits colorectal cancer metastasis and reverses immunotherapy resistance by suppressing angiogenesis

doi: 10.1186/s13046-025-03422-7

Figure Lengend Snippet: Slc9a2 effectively reverses immune resistance in colorectal cancer by inhibiting angiogenesis. ( A ) Flowchart for the establishment of the MC38-R cell line demonstrating resistance to immunotherapy. MC38 cells that developed acquired resistance to immunotherapy were selected under low-dose PD-1 antibody pressure and designated as MC38-R. ( B ) Subcutaneous tumor models in C57BL/6 mice were constructed using both MC38-S (sensitive) and MC38-R cell lines ( n = 4/group). Subsequent intraperitoneal injection of PD-1 monoclonal antibody was conducted to evaluate the efficacy of immunotherapy based on tumor weight. ( C ) Three mice from each group depicted in Fig. 7B were selected for H&E staining, as well as IHC staining for CD8 and CD31, followed by statistical analysis. ( D ) MC38-R cells were employed to establish subcutaneous tumor models in C57BL/6 mice. Upon reaching a tumor volume of 100 mm³, an SLC9A2 overexpression plasmid was intratumorally injected, accompanied by intraperitoneal administration of anti-PD-1 and oral administration of Bevacizumab. ( E-G ) Following four weeks of MC38-R subcutaneous tumor implantation, the mice were sacrificed, and tumors were dissected for photography and weight measurement ( G ). The volume changes of tumor growth were analyzed for each group ( E ), alongside assessments of body weight variations in the mice ( F ). ( H ) VEGFA levels in tumor tissues from each group of mice were quantified using ELISA, with the control group mean normalized to 1 for statistical analysis and graphical representation. ( I-M ) Tumor tissues depicted in Fig. 7E were embedded in paraffin and subjected to H&E staining ( I ), as well as CD31 ( J ) and CD8 ( K ) immunohistochemistry. Additionally, TUNEL ( L ) and EdU ( M ) staining were performed to assess tumor proliferation and apoptosis. ( N ) Granzyme B (left) and IFNγ (right) levels in tumor tissues from each group of mice were quantified using ELISA, with the control group mean normalized to 1 for statistical analysis and graphical representation. ( O ) Map of the slc9a2 pcDNA plasmid and validation of the transfection efficiency in MC38. ( P ) Western blot analysis was conducted to determine the levels of VEGFA and p-STAT3 Y705 . Data in bar graphs indicate mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s., not significant. Student’s t test ( B , C ), multi-group analysis of variance ( G , H , J , K , L , M , N )

Article Snippet: The p-STAT3 Y705 inhibitor Stattic (MCE, HY-13818) and the JAK1/2 inhibitor Ruxolitinib (MCE, HY-50856) were utilized in this study.

Techniques: Construct, Injection, Staining, Immunohistochemistry, Over Expression, Plasmid Preparation, Tumor Implantation, Enzyme-linked Immunosorbent Assay, Control, TUNEL Assay, Biomarker Discovery, Transfection, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Tumor suppressor SLC9A2 inhibits colorectal cancer metastasis and reverses immunotherapy resistance by suppressing angiogenesis

doi: 10.1186/s13046-025-03422-7

Figure Lengend Snippet: Ruxolitinib Augments the Effectiveness of Anti-PD-1 Immunotherapy against CRC. ( A ) MC38-R cells were used to establish subcutaneous tumor models in C57BL/6 mice. Starting on day 10, mice received intraperitoneal injections of anti-PD1, followed by oral administration of ruxolitinib starting on day 13. ( B-D ) Mice were sacrificed two weeks later, and tumors were harvested for photography and weight measurement ( B ). Tumor volume changes were analyzed for each group ( D ), along with assessments of body weight variations in the mice ( C ). ( E ) Western blot analysis was conducted to determine the levels of VEGFA and p-STAT3 Y705 in the tumor tissues. ( F-G ) Levels of VEGFA ( F ), IFNγ, and Granzyme B ( G ) in tumor tissues from each group of mice were quantified using ELISA. The mean of the control group was normalized to 1 for statistical analysis and graphical representation. ( H ) Tumor tissues were embedded in paraffin and subjected to H&E staining, as well as immunohistochemical analysis for CD8, CD31, Ki-67 and TUNEL staining. ( I ) Diagram illustrating the proposed mechanism by which SLC9A2 inhibits immune evasion in CRC. Data in bar graphs indicate mean ± SEM, and data were analyzed using multi-group analysis of variance. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s., not significant

Article Snippet: The p-STAT3 Y705 inhibitor Stattic (MCE, HY-13818) and the JAK1/2 inhibitor Ruxolitinib (MCE, HY-50856) were utilized in this study.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Control, Staining, Immunohistochemical staining, TUNEL Assay

Journal: Nature Communications

Article Title: Macrophage STING signaling promotes fibrosis in benign airway stenosis via an IL6-STAT3 pathway

doi: 10.1038/s41467-024-55170-5

Figure Lengend Snippet: a GSEA analysis showing TOP5 enriched hallmark pathways in Granulation group compared with Normal group. b GSEA analysis show IL6 jak STAT3 pathway enrichment in Granulation group. c Box Plot showing the JAK-STAT pathway score in all clusters in Normal group and Granulation group: B cells ( n = 1434), basal cells(n = 10982), ciliated cells ( n = 1317), endothelial cells ( n = 7241), epithelial cells ( n = 5123), fibroblasts ( n = 6808), mast cells ( n = 475), macrophages ( n = 5625), neutrophils ( n = 10178), plasma cells ( n = 1249), proliferating cells ( n = 824), SMCs ( n = 3608), T cells ( n = 3500) and Type II alveolar cells ( n = 222). n represents the number of cells in cluster. d Representative images of P-STAT3 and P-STING immunohistochemical staining. ( n = 3 sample per group). e Heatmap illustrates ligand-receptor interactions between Macrophages, T cells, neutrophils, and fibroblasts. f – r Mice were divided into three groups: control, BAS + PBS and BAS + Clodronate Liposomes. f Schematic of Clodronate Liposomes administration in mice. g Representative images of H&E staining in different groups at days 7 and 14. h Quantitative analysis of LP thickness in different groups. ( n = 4 mice per group). i Micro CT scans of different groups in Horizontal, Coronal and Sagittal plane. j SYNPASE 3D reconstruction of mouse trachea. Red arrows indicating the stenosis site in ( i – k ). The CT diagram of the area of tracheal stenosis measured by SYNPASE 3D and quantitative analysis of area in different groups(n = 4 mice per group). l Representative images of Masson staining in different groups. m Quantitative analysis of collagen area in different groups ( n = 4 mice per group). n , o Representative tracheal immunofluorescence images and mean fluorescence intensity of αSMA(red) and COL1(green) in different groups ( n = 3 mice per group). p GSEA analysis show IL6 jak STAT3 pathway enrichment in BAS + PBS group. q , r Representative tracheal immunofluorescence images and mean fluorescence intensity of P-STAT3(green) and IL6(red) in different groups ( n = 3 mice per group). Scale bars in ( d , g , n , q ) indicates 200 μm and 50 μm. Scale bars in ( l ) indicate 50 μm. In ( c ), box bounds shows 25th and 75th percentiles, whiskers shows 25th percentiles minus 1.5*IQR to 75th percentiles plus 1.5*IQR and box center shows the median. Data are presented as the mean ± SEM. One-way ANOVA analysis followed by Tukey post hoc multi-comparison test was used in ( h , k , m , o , r ). One-way ANOVA analysis followed by Duncan multiple- range test was used in c . Source data are provided as a Source Data file.

Article Snippet: For inhibit P-STAT3 in mice, the mice were given 10 mg/kg Stattic(#HY-13818, Med-Chem Express, USA) dissolved in 10% DMSO + 40% PEG300(#HY-Y0873, Med-Chem Express, USA) + 5%Tween-80(#HY-Y1891, Med-Chem Express, USA) + 45% saline by intraperitoneal injection on 2 h before modeling, days 2, 4 and 6 after modeling.

Techniques: Immunohistochemical staining, Staining, Control, Liposomes, Micro-CT, Immunofluorescence, Fluorescence, Comparison

Journal: Nature Communications

Article Title: Macrophage STING signaling promotes fibrosis in benign airway stenosis via an IL6-STAT3 pathway

doi: 10.1038/s41467-024-55170-5

Figure Lengend Snippet: a Gene ontology (GO) functional enrichment analysis showed that the enriched DNA damage and repair genes in the BAS group at 24 h after tracheal injury compare to control group ( n = 3 mice per group). b Representative images of H&E staining and immunofluorescence of γH2AX(green) in mouse trachea( n = 3 mice per group). Scale bars indicate 50 μm in H&E staining images, 50 μm and 10 μm in immunofluorescence images. c Representative tracheal immunofluorescence images of dsDNA(pink), F4/80(red), P-STING (green), cGAS(yellow) in mouse trachea ( n = 3 mice per group). Scale bars indicate 200 μm and 50 μm. d Schematic of use DECS to stimulate BMDMs. e Quantification analysis of dsDNA in cell culture supernatant in control group and DECS group ( n = 4 independent experiments). f Representative images of AGAR gel electrophoresis of cell supernatants from control and DECS groups ( n = 3 independent experiments). g , h Protein levels of phosphorylated STING in BMDM group stimulated with or without DECS, n = 3 independent experiments. i Schematic of BMDMs from WT mice stimulated by transfection of mouse tracheal DNA by lipo3000. j , k Protein levels of phosphorylated STING in the BMDM group stimulated with or without DNA ( n = 3 independent experiments). l Representative immunofluorescence images of P-STING(green) in BMDM, Scale bars indicate 50 μm. m Quantitative analysis of relative Mean fluorescence intensity of P-STING ( n = 5 independent experiments). o Protein levels of IL6 in the supernatant of BMDM group stimulated with or without DNA ( n = 4 independent experiments). p mRNA levels of IL6 in control group, DNA stimulated group and DNA stimulated group treated with C176 ( n = 4 independent experiments). q , r Representative tracheal immunofluorescence images of and quantitative analysis of mean fluorescence intensity of P-STAT3(green) and IL6(red) in the different groups ( n = 3 mice per group). Scale bars indicate 200 μm and 20 μm. s Schematic diagram depicting in-vitro co-culture model. t , u Representative immunofluorescence images of and quantitative analysis of mean fluorescence intensity of P-STAT3(red) and αSMA(green) in fibroblasts ( n = 3 independent experiments). Scale bars indicate 10 μm. Data are presented as the mean ± SEM. A two-sided student’s T-test was used in ( e , h , k , o ). One-way ANOVA analysis followed by Tukey post hoc multi-comparison test was used in ( m , p , r , u ). Source data are provided as a Source Data file.

Article Snippet: For inhibit P-STAT3 in mice, the mice were given 10 mg/kg Stattic(#HY-13818, Med-Chem Express, USA) dissolved in 10% DMSO + 40% PEG300(#HY-Y0873, Med-Chem Express, USA) + 5%Tween-80(#HY-Y1891, Med-Chem Express, USA) + 45% saline by intraperitoneal injection on 2 h before modeling, days 2, 4 and 6 after modeling.

Techniques: Functional Assay, Control, Staining, Immunofluorescence, Cell Culture, Nucleic Acid Electrophoresis, Transfection, Fluorescence, In Vitro, Co-Culture Assay, Comparison

Journal: Nature Communications

Article Title: Macrophage STING signaling promotes fibrosis in benign airway stenosis via an IL6-STAT3 pathway

doi: 10.1038/s41467-024-55170-5

Figure Lengend Snippet: Upon injury, dsDNA released from the tracheal epithelium activates the STING pathway in macrophages, leading to the release of the inflammatory cytokine IL-6. IL-6 then activates STAT3 signaling in fibroblasts, contributing to the fibrosis of granulation tissue. Treatment with the cGAS inhibitor RU.521, the STING inhibitor C176, or IL-6 neutralizing antibody alleviates granulation tissue fibrosis. Additionally, depletion of macrophages using bisphosphonate liposomes reduces the extent of fibrosis in the granulation tissue.

Article Snippet: For inhibit P-STAT3 in mice, the mice were given 10 mg/kg Stattic(#HY-13818, Med-Chem Express, USA) dissolved in 10% DMSO + 40% PEG300(#HY-Y0873, Med-Chem Express, USA) + 5%Tween-80(#HY-Y1891, Med-Chem Express, USA) + 45% saline by intraperitoneal injection on 2 h before modeling, days 2, 4 and 6 after modeling.

Techniques: Liposomes

Journal: Communications Biology

Article Title: Decidual natural killer cells dysfunction is caused by IDO downregulation in dMDSCs with Toxoplasma gondii infection

doi: 10.1038/s42003-024-06365-5

Figure Lengend Snippet: a The expression of p52, RelB and IDO in purified human dMDSCs of uninfected and infected groups were examined by qPCR (data represent the mean ± SD from five independent experiments). b The protein levels of IDO, STAT3, p-STAT3 (Y705), IKKα, p-IKKα, p-p100 in purified human dMDSCs, p52 and RelB in nucleus of dMDSCs of uninfected and infected groups were determined by western blot. c The statistical analysis of IDO, p-STAT3 (Y705), p-IKKα, p-p100, p52 and RelB in protein levels from infected groups and uninfected human dMDSCs by western blot. (data represent the mean ± SD from three independent experiments). d The expression levels of p-STAT3 in mouse dMDSCs were examined by flow cytometry in uninfected ( n = 7) and infected groups ( n = 8) and the results were analyzed by statistical analysis. Data are presented as the mean ± SD, * P < 0.05, ** P < 0.01, human samples in each group assayed individually by the paired t -test and mice samples in each group assayed individually by the unpaired t -test. MFI: mean fluorescence intensity.

Article Snippet: To investigate if IDO expression was regulated by the STAT3/p52-RelB pathway after T. gondii infection, the p-STAT3 inhibitor (stattic; MCE, China) and p52-RelB nuclear translocation inhibitor (SN52; MCE, China) were separately introduced to human dMDSCs.

Techniques: Expressing, Purification, Infection, Western Blot, Flow Cytometry, Fluorescence

Journal: Communications Biology

Article Title: Decidual natural killer cells dysfunction is caused by IDO downregulation in dMDSCs with Toxoplasma gondii infection

doi: 10.1038/s42003-024-06365-5

Figure Lengend Snippet: a The mRNA levels of IDO in human dMDSCs of infected group, infected with static treated group and infected with SN52 treated group were detected by qPCR (data represent the mean ± SD from three independent experiments by one-way ANOVA). b The expression changes of STAT3, p-STAT3 (Y705), IKKα, p-IKKα, p-p100, IDO and the expression of p52 and RelB of nucleus in purifed human dMDSCs of the uninfected, infected and infected with static treated groups were determined by western blot (data represent the mean ± SD from three independent experiments by one-way ANOVA). c The expression changes of IDO in human dMDSCs and the expression of p52 and RelB in nucleus of dMDSCs from the infected and infected with SN52 treated groups were determined by western blot (data represent the mean ± SD from three independent experiments by the paired t -test). d The IDO expression in mouse dMDSCs of infected and infected with JSI-124 groups were examined by flow cytometry ( n = 6). Data are presented as the mean ± SD, * P < 0.05, ** P < 0.01, unpaired t -test. MFI mean fluorescence intensity.

Article Snippet: To investigate if IDO expression was regulated by the STAT3/p52-RelB pathway after T. gondii infection, the p-STAT3 inhibitor (stattic; MCE, China) and p52-RelB nuclear translocation inhibitor (SN52; MCE, China) were separately introduced to human dMDSCs.

Techniques: Infection, Expressing, Western Blot, Flow Cytometry, Fluorescence

Journal: Communications Biology

Article Title: Decidual natural killer cells dysfunction is caused by IDO downregulation in dMDSCs with Toxoplasma gondii infection

doi: 10.1038/s42003-024-06365-5

Figure Lengend Snippet: T. gondii infection upregulated the transcriptional levels of IDO in dMDSCs through STAT3/p52-RelB pathway. But IDO protein expression was decreased due to the degradation induced by the increase of SOCS3 by infection, which in turn the reduction of IDO metabolite kynurenine further downregulated TGF-β and IL-10 expression in dNK cells through Kyn/AhR/SP1 signaling pathway, ultimately leading to the dysfunction of dNK and the disorders of maternal-fetal tolerance.

Article Snippet: To investigate if IDO expression was regulated by the STAT3/p52-RelB pathway after T. gondii infection, the p-STAT3 inhibitor (stattic; MCE, China) and p52-RelB nuclear translocation inhibitor (SN52; MCE, China) were separately introduced to human dMDSCs.

Techniques: Infection, Expressing