stat3 inhibitor ag490  (MedChemExpress)

Journal: Breast Cancer Research : BCR

Article Title: Modulating phosphodiesterase-5 activity to suppress the immunosuppressive mechanisms of myeloid-derived suppressor cells in breast cancer

doi: 10.1186/s13058-025-02131-5

Figure Lengend Snippet: Tadalafil disrupted the immunosuppressive ability of MDSCs by inhibiting STAT3 transcription. A RT-qPCR analysis of the expression of the BM-MDSCs stat3 . B Western blot analysis was used to detect STAT3 expression in BM-MDSCs after tadalafil treatment. C Flow cytometry representative plot of the effect of tadalafil treatment combined with administration of the STAT3 inhibitor on the expression of arginase 1, a functional marker of MDSCs, induced in vitro. D Transwell co-culture plate design. E Representative images and quantification of heme staining in transwell chambers. Data represents the mean ± SD ( n ≥ 3), **P < 0.01 and **P < 0.001 versus vehicle

Article Snippet: Tadalafil (HY-90009 A), STAT3 inhibitor (Tyrphostin AG490, HY-12000) and PARP-1 inhibitor (AG14361, HY-12032) were purchased from MedChem Express (New Jersey, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Flow Cytometry, Functional Assay, Marker, In Vitro, Co-Culture Assay, Staining

Journal: Breast Cancer Research : BCR

Article Title: Modulating phosphodiesterase-5 activity to suppress the immunosuppressive mechanisms of myeloid-derived suppressor cells in breast cancer

doi: 10.1186/s13058-025-02131-5

Figure Lengend Snippet: Tadalafil regulates STAT3 transcription through PARP1. A GSEA analysis revealed significant alterations in NAD-related signaling pathways following tadalafil intervention in BM-MDSCs. B Heatmap displaying the expression patterns of genes associated with NAD-related signaling pathways. C Flow cytometry gating strategy and analysis of PARP1 median fluorescence intensity in BM-MDSCs. D Western blot analysis of cGKI and PARP1 expression in MDSC cells after tadalafil treatment. E The expression level of PARP1 mRNA in BM-MDSCs. F Immunofluorescence staining assessing PARP1 expression in BM-MDSCs post-tadalafil treatment. G Western blot detection of PARP1, STAT3, and phosphorylated STAT3 (p-STAT3) levels in cytoplasmic and nuclear fractions of BM-MDSCs upon tadalafil intervention. H Western blot analysis of STAT3 and p-STAT3 expression changes in BM-MDSCs treated with tadalafil in the presence of a PARP1 inhibitor. I Flow cytometry gating strategy and analysis of p-STAT3 median fluorescence intensity in BM-MDSCs. J Genotyping verification of PARP1 knockout mice. K Western blot confirmation of PARP1 knockout efficiency. L Western blot evaluation of Arg1, STAT3, and p-STAT3 expression in BM-MDSCs from PARP1 knockout mice following tadalafil intervention. Data represents the mean ± SD ( n ≥ 3), *P < 0.05, **P < 0.01 and **P < 0.001 vs. vehicle

Article Snippet: Tadalafil (HY-90009 A), STAT3 inhibitor (Tyrphostin AG490, HY-12000) and PARP-1 inhibitor (AG14361, HY-12032) were purchased from MedChem Express (New Jersey, USA).

Techniques: Protein-Protein interactions, Expressing, Flow Cytometry, Fluorescence, Western Blot, Immunofluorescence, Staining, Knock-Out

Journal: Cell Biology and Toxicology

Article Title: METTL3, m6A modification, and EGR1: interplay affecting myocardial I/R injury outcomes

doi: 10.1007/s10565-024-09937-7

Figure Lengend Snippet: The effect of EGR1 on mitophagy through the regulation of the JAK2/STAT3 pathway. Note: ( A ) Western blot analysis of the expression and quantification of JAK2/STAT3 pathway-related proteins in cardiomyocytes from different treatment groups; ( B ) Schematic diagram showing the treatment of AG490 after shEGR1 transfection in the H/R damage model; ( C ) Western blot analysis of the expression and quantification of JAK2/STAT3 pathway-related proteins in cardiomyocytes from different treatment groups; ( D ) TEM to observe the morphology of cardiomyocyte mitochondria in each group, with a scale bar of 500 nm and arrows indicating mitochondria; ( E ) Representative immunofluorescence images showing the co-localization of GFP-LC3B (green) and mitochondria (MTR-Red, red) in cardiomyocytes from different treatment groups, with a scale bar of 25 μm, and quantification of the number of co-localized spots between GFP-LC3B and mitochondria, with DAPI (blue) indicating the nucleus; ( F ) Representative immunofluorescence images showing the co-localization of MTR-Green (green) and lysosomes (LTR, red) in cardiomyocytes from different treatment groups, with a scale bar of 50 μm, and quantification of the number of co-localized spots between lysosomes and mitochondria in each cell; ( G ) Western blot analysis of the expression and quantification of mitophagy-related proteins in cardiomyocytes from different treatment groups; ( H ) Assessment of cell viability of cardiomyocytes in different treatment groups using the CCK-8 method; ( I ) Measurement of the levels of cTnI and CK-MB in the supernatant of cardiomyocytes in different treatment groups using the ELISA method. * indicates a significant difference between two groups with P < 0.05, ** indicates a significant difference between two groups with P < 0.01, *** indicates a significant difference between two groups with P < 0.001, **** indicates a significant difference between two groups with P < 0.0001. All experiments were repeated three times

Article Snippet: Furthermore, the H/R+shEGR1+3-MA and H/R+shEGR1+AG490 groups were transfected with shEGR1 and treated with the mitophagy inhibitor 3-Methyladenine (3-MA, 5 mM; MCE, HY-19312) or the JAK2/STAT3 pathway-specific inhibitor AG490 (50 μM; MCE, HY-12000) during H/R model construction for 36 hours (Chen et al. ; Zeng et al. ; Yin et al. ).

Techniques: Western Blot, Expressing, Transfection, Immunofluorescence, CCK-8 Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Biology and Toxicology

Article Title: METTL3, m6A modification, and EGR1: interplay affecting myocardial I/R injury outcomes

doi: 10.1007/s10565-024-09937-7

Figure Lengend Snippet: The Impact of EGR1/JAK2/STAT3 axis-mediated mitophagy dysfunction on pyroptosis. Note: ( A ) Ultrastructural morphology of cardiomyocytes observed under SEM. Scale bar=10 μm; ( B ) Representative images of TUNEL staining in cardiomyocytes from each group (Scale bar=50 μm) and the percentage of TUNEL-positive cells; ( C ) LDH release results in cardiomyocytes from each group measured by ELISA; ( D ) Expression and quantification of pyroptosis-related proteins in myocardial cells from each group detected by Western blot; ( E ) Levels of IL1β and IL18 in the supernatant of cardiomyocytes from each group measured by ELISA; ( F ) Ultrastructural morphology of cardiomyocytes observed under SEM. Scale bar=10 μm; ( G ) Representative images of TUNEL staining in cardiomyocytes from each group (Scale bar=50 μm) and the percentage of TUNEL-positive cells; ( H ) LDH release results in cardiomyocytes from each group measured by ELISA; ( I ) Expression and quantification of pyroptosis-related proteins in myocardial cells from each group detected by Western blot; ( J ) Levels of IL1β and IL18 in the supernatant of cardiomyocytes from each group measured by ELISA; C1-Cas1: Cleaved-Caspase 1; * indicates p < 0.05 compared to the control group, ** indicates p < 0.01, *** indicates p < 0.001, **** indicates p < 0.0001. All experiments were repeated three times

Article Snippet: Furthermore, the H/R+shEGR1+3-MA and H/R+shEGR1+AG490 groups were transfected with shEGR1 and treated with the mitophagy inhibitor 3-Methyladenine (3-MA, 5 mM; MCE, HY-19312) or the JAK2/STAT3 pathway-specific inhibitor AG490 (50 μM; MCE, HY-12000) during H/R model construction for 36 hours (Chen et al. ; Zeng et al. ; Yin et al. ).

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

Journal: Cell Biology and Toxicology

Article Title: METTL3, m6A modification, and EGR1: interplay affecting myocardial I/R injury outcomes

doi: 10.1007/s10565-024-09937-7

Figure Lengend Snippet: The effect of METTL3 on the characterization of I/R mice through EGR1/JAK2/STAT3 pathway. Note: ( A ) Expression of METTL3 and EGR1 in cardiac tissue of different groups of mice (n=8) as measured by RT-qPCR; ( B ) Protein expression and quantification of METTL3 and EGR1 in cardiac tissue of different groups of mice (n=8) as determined by Western blot; ( C ) Expression and quantification of JAK2/STAT3 pathway-related proteins in cardiac tissue of different groups of mice (n=8) as measured by Western blot; ( D ) Cardiac ultrasound evaluation of heart function-related indices in different groups of mice (n=6); ( E ) Representative images of Evans blue/TTC double staining in cardiac tissue of different groups of mice (n=8), with blue regions representing normal cardiac tissue, red regions representing ischemic myocardium (AAR), and white regions representing the infarct area (INF) of cardiac tissue. Quantification of INF/AAR and AAR/LV percentages, where LV represents the left ventricle; ( F ) Representative images of HE-stained cardiac tissue in different groups of mice (n=8), Scale bar=50 μm; ( G ) Detection of cTnI and CK-MB levels in serum of different groups of mice (n=8) using ELISA; * indicates a significant difference ( p < 0.05) between two groups, ** indicates a significant difference ( p < 0.01) between two groups, *** indicates a highly significant difference ( p < 0.001) between two groups, **** indicates an extremely significant difference ( p < 0.0001) between two groups

Article Snippet: Furthermore, the H/R+shEGR1+3-MA and H/R+shEGR1+AG490 groups were transfected with shEGR1 and treated with the mitophagy inhibitor 3-Methyladenine (3-MA, 5 mM; MCE, HY-19312) or the JAK2/STAT3 pathway-specific inhibitor AG490 (50 μM; MCE, HY-12000) during H/R model construction for 36 hours (Chen et al. ; Zeng et al. ; Yin et al. ).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Double Staining, Staining, Enzyme-linked Immunosorbent Assay