stat3 phosphorylation inhibitor stattic  (MedChemExpress)

Journal: Hepatology (Baltimore, Md.)

Article Title: Hepatic GDP-fucose transporter SLC35C1 attenuates cholestatic liver injury and inflammation by inducing CEACAM1 N153 fucosylation

doi: 10.1097/HEP.0000000000001003

Figure Lengend Snippet: Conjugated bile acids enhance the binding of activated STAT3 to the SLC35C1 promoter to directly induce SLC35C1 expression in cholestasis. (A) A diagram illustrated the potential STAT3 binding sites in human SLC35C1 proximal promoter and the reporter constructs. (B) Treatment with TCA-induced STAT3 activation and SLC35C1 expression in PLC/PRF/5- ASBT cells, which was mitigated by treatment with a STAT3-specific inhibitor (APTSTAT3-9R). n=3, *p <0.05 vs. the DMSO group; # p <0.05 vs. the cells treated with 100 μM TCA alone. (C) Representative western blot detecting leakage of endoplasmic reticulum protein Grp78 into the cytosol in mouse hepatocytes after a 24-hour treatment with 100 μM TCA. n=3, *p <0.05 vs. the DMSO group. (D) PLC/PRF/5- ASBT cells were transiently transfected with 4 different truncated SLC35C1 promoters and treated with or without TCA. Their luciferase activities were measured. (E) PLC/PRF/5- ASBT cells were co - transfected with 4 different truncated SLC35C1 promoters and the plasmid for STAT3 overexpression or control, with or without TCA treatment. (F) PLC/PRF/5- ASBT cells were co-transfected with WT or mutant SLC35C1 promoter region and the plasmid for STAT3 overexpression or control, with or without TCA treatment. *p <0.05. (G) ChIP assays revealed that the TCA-induced STAT3 binding to the SLC35C1 -827 was abrogated by treatment with APTSTAT3-9R in PLC/PRF/5- ASBT cells. *p <0.05 vs. the cells treated without TCA; # p <0.05 vs the cells treated with 100 μM TCA. (H) ChIP assays displayed the binding of activated STAT3 to the SLC35C1 -827 in liver tissues of patient with OC. (I, J) ChIP assays exhibited the binding of activated Stat3 to the Slc35c1 -693 in the livers of BDL mouse model and 1% CA-fed mouse model. Abbreviations: BDL, bile duct ligation; ChIP, chromatin immunoprecipitation; CTR, control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PLC, primary liver carcinoma; PRF, Poliomyelitis Research Foundation; SLC35C1, Solute Carrier Family 35 Member C1; RPF, Poliomyelitis Research Foundation; TCA, taurocholic acid; qPCR, quantitative polymerase chain reaction; WT, wild type.

Article Snippet: APTSTAT3-9R (Cat# S8197), an inhibitor of STAT3 phosphorylation, was from Selleck Chem (Houston, TX).

Techniques: Binding Assay, Expressing, Construct, Activation Assay, Western Blot, Transfection, Luciferase, Plasmid Preparation, Over Expression, Control, Mutagenesis, Ligation, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction

Journal: Hepatology (Baltimore, Md.)

Article Title: Hepatic GDP-fucose transporter SLC35C1 attenuates cholestatic liver injury and inflammation by inducing CEACAM1 N153 fucosylation

doi: 10.1097/HEP.0000000000001003

Figure Lengend Snippet: A schematic diagram illustrates a novel negative feedback loop against cholestatic liver injury and inflammation. Initially, the accumulated BAs in the liver trigger inflammatory responses and cause liver injury during the process of cholestasis. Meanwhile, BAs also activate the STAT3 signaling and enhance the binding of activated STAT3 to the SLC35C1 promoter to upregulate SLC35C1 expression in hepatocytes. Finally, elevated SLC35C1 expression facilitates CEACAM1 fucosylation, which in turn suppresses CCL2 and CXCL2 expression to inhibit liver inflammation during the process of cholestasis. Abbreviation: SLC35C1, Solute Carrier Family 35 Member C1.

Article Snippet: APTSTAT3-9R (Cat# S8197), an inhibitor of STAT3 phosphorylation, was from Selleck Chem (Houston, TX).

Techniques: Binding Assay, Expressing

Journal: Brain Sciences

Article Title: Minocycline Ameliorates Staphylococcus aureus -Induced Neuroinflammation and Anxiety-like Behaviors by Regulating the TLR2 and STAT3 Pathways in Microglia

doi: 10.3390/brainsci15020128

Figure Lengend Snippet: Effects of minocycline on STAT3 activation and GLS1 expression in LTA-induced microglial cells. ( A , B ) The expression levels of p-STAT3, STAT3, and GLS1 in the BV2 and N9 cells were analyzed via Western blot analysis. ( C , D ) The quantification of the expression levels of p-STAT3/STAT3 and GLS1 in BV2 and N9 cells. The data were normalized according to GAPDH. The mean ± SEM of three separate experiments is represented for all data that have error bars. Compared to the control group, * p < 0.05; ** p < 0.01; *** p < 0.001. Compared to the LTA group, # p < 0.05; ## p < 0.01; ### p < 0.001. Control, saline-treated group; LTA, cells pretreated with saline and then stimulated with LTA; Mino100 + LTA, cells pretreated with 100 μmol/L minocycline and then stimulated with LTA; Mino200 + LTA, cells pretreated with 200 μmol/L minocycline and then stimulated with LTA.

Article Snippet: The STAT3 phosphorylation (at Y705 and S727) inhibitor Stattic was purchased from MCE (Princeton, NJ, USA).

Techniques: Activation Assay, Expressing, Western Blot, Control, Saline

Journal: Brain Sciences

Article Title: Minocycline Ameliorates Staphylococcus aureus -Induced Neuroinflammation and Anxiety-like Behaviors by Regulating the TLR2 and STAT3 Pathways in Microglia

doi: 10.3390/brainsci15020128

Figure Lengend Snippet: The inhibition of p-STAT3 reduces the proinflammatory cytokines and GLS1 levels in LTA-induced microglial cells. ( A , B ) The TNF-α, IL-6, and IL-10 secreted by the BV2 and N9 cells were assessed via ELISA. ( C , D ) The expression levels of TLR2, TNF-α, IL-6, GLS1, p-STAT3, and STAT3 in the BV2 and N9 cells were analyzed via Western blot analysis. ( E , F ) The quantification of the expression levels of p-STAT3/STAT3, TLR2, TNF-α, IL-6, and GLS1 in the BV2 and N9 cells. The data were normalized according to GAPDH. The mean ± SEM of three separate experiments is represented for all data that have error bars. Compared to the control group, * p < 0.05; ** p < 0.01; *** p < 0.001. Compared to the LTA group, # p < 0.05; ## p < 0.01; ### p < 0.001. Control, saline-treated group; LTA, cells pretreated with saline and then stimulated with LTA; Stattic, cells pretreated with 10 μmol/L Stattic and then treated with saline; Stattic + LTA, cells pretreated with 10 μmol/L Stattic and then stimulated with LTA.

Article Snippet: The STAT3 phosphorylation (at Y705 and S727) inhibitor Stattic was purchased from MCE (Princeton, NJ, USA).

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Control, Saline

Journal: Brain Sciences

Article Title: Minocycline Ameliorates Staphylococcus aureus -Induced Neuroinflammation and Anxiety-like Behaviors by Regulating the TLR2 and STAT3 Pathways in Microglia

doi: 10.3390/brainsci15020128

Figure Lengend Snippet: Effects of minocycline on S. aureus -induced neuroinflammation in vivo. ( A ) The GLS1, p-STAT3, and STAT3 expression levels in the mPFC of two representative mice were analyzed via Western blot analysis. ( B ) The expression levels of p-STAT3/STAT3 and GLS1 in the mPFC were quantified. The data were normalized according to GAPDH. Data are presented as mean ± SEM, n = 4. ( C ) The TLR2, TNF-α, and IL-6 protein expression levels in the mPFC of two representative mice were analyzed via Western blot analysis. ( D ) The expression levels of TLR2, TNF-α, and IL-6 in the mPFC were quantified. The data were normalized according to GAPDH. Data are presented as mean ± SEM, n = 4. ( E ) Schematic diagram showing the mPFC area (red box) of the mice analyzed via immunofluorescent staining. ( F ) Representative fluorescence micrographs showing the number of microglia in the mPFC in the various groups (IBA1, red; DAPI, blue); scale bar = 100 μm. ( G ) A quantitative analysis of the number of IBA1 + cells in the mPFC. Data are presented as mean ± SEM, n = 3. USA300 group compared to the control group, * p < 0.05. Mino + USA300 group compared to the USA300 group, # p < 0.05; ## p < 0.01. Control, saline-treated group; Mino, mice pretreated with minocycline and then challenged with the saline group; USA300, mice pretreated with saline and then challenged with the USA300 group; Mino + USA300, mice pretreated with minocycline and then challenged with the USA300 group.

Article Snippet: The STAT3 phosphorylation (at Y705 and S727) inhibitor Stattic was purchased from MCE (Princeton, NJ, USA).

Techniques: In Vivo, Expressing, Western Blot, Staining, Fluorescence, Control, Saline