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inhibitors mln120b  (MedChemExpress)
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MedChemExpress inhibitors mln120b
The activation of the STAT3 signaling pathway by Lp is mediated by the RIG-I-like signaling pathway. ( A ) Pan-JAK inhibitor Pyridone 6 inhibits the activation of STAT3 induced by LCMV Lp. HEK-293T cells in a well of a 24-well plate were co-transfected with 480 ng of pCAGGs-LCMV-Lp and 20 ng STAT3 reporter plasmids. At 6 h post transfection, cells were treated with the indicated concentration of Pyridone 6. At 36 h post transfection, the cells were lysed to measure the STAT3 reporter activity using a luciferase assay. ( B ) IKK-β inhibitor <t>MLN120B</t> inhibits the activation of STAT3 induced by LCMV Lp. HEK-293T cells in a well of a 24-well plate were co-transfected with 480 ng of pCAGGs-LCMV-Lp and 20 ng of STAT3 reporter plasmids. At 6 h post transfection, cells were treated with the indicated concentration of MLN120B. At 36 h post transfection, the cells were lysed to measure the STAT3 reporter activity using a luciferase assay. ( C ) TBK1 and IKK-ε inhibitor Amlexanox inhibits the activation of STAT3 induced by LCMV Lp. HEK-293T cells in a well of a 24-well plate were co-transfected with 480 ng of pCAGGs-LCMV-Lp and 20 ng of STAT3 reporter plasmids. At 6 h post transfection, cells were treated with the indicated concentration of Amlexanox. At 36 h post transfection, the cells were lysed to measure the STAT3 reporter activity using a luciferase assay. ( D – F ) The effect of Pyridone 6, MLN120B, and Amlexanox on the viability of HEK-293T cells. ( G – I ) The effect of the knockdown of RIG-I, MDA5, or MAVS on the activation of STAT3 mediated by LCMV Lp. HEK-293T cells in a well of a 24-well plate were trans-transfected with the indicated siRNAs. At 24 h post transfection, cells were transfected with 20 ng of STAT3 reporter plasmid and 480 ng of pCAGGs-LCMV-Lp. Thirty-six hours later, the cells were collected. Intracellular RNA was extracted and the intracellular level of RIG-I, MDA5, or MAVS was measured using qRT-PCR (left). For luciferase assay, cells were collected and the intracellular level of STAT3 reporter activity was measured (right). A representative from at least three independent experiments is shown. Graphs show mean ± SD. ( n = 3, A – I ). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, unpaired Student’s t -test.
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The activation of the STAT3 signaling pathway by Lp is mediated by the RIG-I-like signaling pathway. ( A ) Pan-JAK inhibitor Pyridone 6 inhibits the activation of STAT3 induced by LCMV Lp. HEK-293T cells in a well of a 24-well plate were co-transfected with 480 ng of pCAGGs-LCMV-Lp and 20 ng STAT3 reporter plasmids. At 6 h post transfection, cells were treated with the indicated concentration of Pyridone 6. At 36 h post transfection, the cells were lysed to measure the STAT3 reporter activity using a luciferase assay. ( B ) IKK-β inhibitor MLN120B inhibits the activation of STAT3 induced by LCMV Lp. HEK-293T cells in a well of a 24-well plate were co-transfected with 480 ng of pCAGGs-LCMV-Lp and 20 ng of STAT3 reporter plasmids. At 6 h post transfection, cells were treated with the indicated concentration of MLN120B. At 36 h post transfection, the cells were lysed to measure the STAT3 reporter activity using a luciferase assay. ( C ) TBK1 and IKK-ε inhibitor Amlexanox inhibits the activation of STAT3 induced by LCMV Lp. HEK-293T cells in a well of a 24-well plate were co-transfected with 480 ng of pCAGGs-LCMV-Lp and 20 ng of STAT3 reporter plasmids. At 6 h post transfection, cells were treated with the indicated concentration of Amlexanox. At 36 h post transfection, the cells were lysed to measure the STAT3 reporter activity using a luciferase assay. ( D – F ) The effect of Pyridone 6, MLN120B, and Amlexanox on the viability of HEK-293T cells. ( G – I ) The effect of the knockdown of RIG-I, MDA5, or MAVS on the activation of STAT3 mediated by LCMV Lp. HEK-293T cells in a well of a 24-well plate were trans-transfected with the indicated siRNAs. At 24 h post transfection, cells were transfected with 20 ng of STAT3 reporter plasmid and 480 ng of pCAGGs-LCMV-Lp. Thirty-six hours later, the cells were collected. Intracellular RNA was extracted and the intracellular level of RIG-I, MDA5, or MAVS was measured using qRT-PCR (left). For luciferase assay, cells were collected and the intracellular level of STAT3 reporter activity was measured (right). A representative from at least three independent experiments is shown. Graphs show mean ± SD. ( n = 3, A – I ). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, unpaired Student’s t -test.

Journal: Viruses

Article Title: Activation of the STAT3 Signaling Pathway by the RNA-Dependent RNA Polymerase Protein of Arenavirus

doi: 10.3390/v13060976

Figure Lengend Snippet: The activation of the STAT3 signaling pathway by Lp is mediated by the RIG-I-like signaling pathway. ( A ) Pan-JAK inhibitor Pyridone 6 inhibits the activation of STAT3 induced by LCMV Lp. HEK-293T cells in a well of a 24-well plate were co-transfected with 480 ng of pCAGGs-LCMV-Lp and 20 ng STAT3 reporter plasmids. At 6 h post transfection, cells were treated with the indicated concentration of Pyridone 6. At 36 h post transfection, the cells were lysed to measure the STAT3 reporter activity using a luciferase assay. ( B ) IKK-β inhibitor MLN120B inhibits the activation of STAT3 induced by LCMV Lp. HEK-293T cells in a well of a 24-well plate were co-transfected with 480 ng of pCAGGs-LCMV-Lp and 20 ng of STAT3 reporter plasmids. At 6 h post transfection, cells were treated with the indicated concentration of MLN120B. At 36 h post transfection, the cells were lysed to measure the STAT3 reporter activity using a luciferase assay. ( C ) TBK1 and IKK-ε inhibitor Amlexanox inhibits the activation of STAT3 induced by LCMV Lp. HEK-293T cells in a well of a 24-well plate were co-transfected with 480 ng of pCAGGs-LCMV-Lp and 20 ng of STAT3 reporter plasmids. At 6 h post transfection, cells were treated with the indicated concentration of Amlexanox. At 36 h post transfection, the cells were lysed to measure the STAT3 reporter activity using a luciferase assay. ( D – F ) The effect of Pyridone 6, MLN120B, and Amlexanox on the viability of HEK-293T cells. ( G – I ) The effect of the knockdown of RIG-I, MDA5, or MAVS on the activation of STAT3 mediated by LCMV Lp. HEK-293T cells in a well of a 24-well plate were trans-transfected with the indicated siRNAs. At 24 h post transfection, cells were transfected with 20 ng of STAT3 reporter plasmid and 480 ng of pCAGGs-LCMV-Lp. Thirty-six hours later, the cells were collected. Intracellular RNA was extracted and the intracellular level of RIG-I, MDA5, or MAVS was measured using qRT-PCR (left). For luciferase assay, cells were collected and the intracellular level of STAT3 reporter activity was measured (right). A representative from at least three independent experiments is shown. Graphs show mean ± SD. ( n = 3, A – I ). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, unpaired Student’s t -test.

Article Snippet: Inhibitors MLN120B (MedChemExpress, Monmouth Junction, NJ, USA, HY-15473), Pyridone 6 (MedChemExpress, HY-14435), and Amlexanox (Selleck Chemicals, Houston, TX, USA, S3648); MagStrep XT beads (IBA Lifesciences, Göttingen, NI, Germany, 2-4090-002) and buffer BXT (IBA Lifesciences, 2-1041-250); recombinant human IL-6 protein (PeproTech, Rocky Hill, NJ, USA, 200-06-5); mouse monoclonal antibodies against GAPDH (Proteintech, Wuhan, Hubei, China, 60004-1-Ig) and V5 (Thermo Fisher Scientific, MA5-15253); rabbit monoclonal antibodies against pY705-STAT3 (Cell Signaling Technology, Danvers, MA, USA, 9145T); rabbit polyclonal antibodies against STAT3 (Proteintech, 10253-2-AP), α-Tubulin (Proteintech, 11224-1-AP), Lamin B1 (Proteintech, 12987-1-AP), and strep-tag (GenScript Biotech, Piscataway, NJ, USA, A00626-40) were purchased from the indicated companies.

Techniques: Activation Assay, Transfection, Concentration Assay, Activity Assay, Luciferase, Knockdown, Plasmid Preparation, Quantitative RT-PCR