jak2 stat3 inhibitor tyrphostin ag490  (MedChemExpress)

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

Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: Network pharmacology and molecular docking of the main components of JQP with the STAT3 protein molecule. A. Venn diagram of candidate target genes of JQP and AS. B. Protein-protein interaction network, the darker the color, the higher the importance. C. Degree screening of the top 20 gene targets. D. Cytoscape 3.7.1 The software draws the active ingredient-target gene network of TCM, in which the yellow square represents the target gene, the green circle represents the traditional Chinese medicine, and the green square represents the active ingredient of the traditional Chinese medicine. E. Bubble chart of KEGG pathway enrichment analysis of candidate AS targets. The size of the dots indicates the number of selected genes, and the color indicates the p value of the enrichment analysis. F. Visualization of molecular docking of active ingredients and core protein targets in the core formulation. (a) Docking diagram of quercetin (MOL000098) and STAT3. (b) Docking diagram of kaempferol (MOL000422) and STAT3. (c) Docking diagram of wogonin (MOL000173) and STAT3. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques: Software, Formulation

Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: Effects of NONHSAT227927.1 on cell viability, inflammatory cytokines and inflammatory pathways.A, Screening of NONHSAT227927.1 small interfering RNA model. B, CCK-8 method was used to detect the cell viability of AS-FLS. C, RT-qPCR was used to detect the expression of NONHSAT227927.1 in AS-FLS. D-G, ELISA was used to detect the levels of IL-6, IL-17, TNF-α, and IL-10 in AS-FLS. H–I, RT-qPCR was used to detect the levels of JAK2 and STAT3 in AS-FLS. J, WB was used to detect Levels of JAK2, STAT, p -JAK2 and p-STAT3 in AS-FLS. K, IF was used to detect the expression of JAK2 and STAT3 in AS-FLS. All experiments were repeated three times.a: ov-NC, b: ov-NONHSAT227927.1, c: si-NC, d: si-NONHSAT227927.1. ***P < 0.001.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques: Small Interfering RNA, CCK-8 Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: NONHSAT227927.1/JAK2/STAT3 combination regulates the activity of AS-FLS and the release of inflammatory factors. A, CCK-8 method was used to detect the cell viability of AS-FLSs. B-E, ELISA method was used to detect the levels of IL-6, IL-17, TNF-α, and IL-10 in AS-FLS. FG, RT-qPCR was used to detect Levels of JAK2 and STAT3 in AS-FLS. H, WB was used to detect the levels of JAK2, STAT, p -JAK2 and p-STAT3 in AS-FLS. I, IF was used to detect the expression of JAK2 and STAT3 in AS-FLS. All experiments were repeated three times. a: FLS, b: AG490, c: ov-NC, d: ov-NONHSAT227927.1, e: AG490 + ov-NONHSAT227927.1.*P < 0.05,**P < 0.01,***P < 0.001.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques: Activity Assay, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing

Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: JQP reversed the effects of NONHSAT227927.1 overexpression on AS-FLS viability, inflammatory factors and JAK2/STAT3 pathway. A, CCK-8 method was used to screen the optimal concentration of JQP-containing serum. B. Use CCK-8 method to detect the cell viability of AS-FLSs. C. RT-qPCR detects the expression of NONHSAT227927.1. D-G used ELISA to detect the levels of IL-6, IL-17, TNF-α, and IL-10. HI, RT-qPCR was used to detect the expression of JAK2 and STAT3. J. WB detects the expression of JAK2, STAT3, p -JAK2 and p-STAT3 proteins. K. Immunofluorescence method detects the expression of p -JAK2 and p-STAT3 proteins. a: FLS, b: NS, c: JQP, d: ov-NC, e: ov-NONHSAT227927.1, f: ov-NONHSAT227927.1 + JQP. The data is displayed in the form of the average value plus or minus the standard deviation. The experiments were conducted thrice.*P < 0.05,**P < 0.01,***P < 0.001.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques: Over Expression, CCK-8 Assay, Concentration Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Standard Deviation

Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: JQP exerts an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques: