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2 protocols using pyridone 6

1

Comprehensive Antibody Inventory for Cellular Signaling

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Antibodies against Jak1, Jak2, Gsk3β, β-catenin, p-Ser9-Gsk3β, p-Ser552-β-catenin, p-Ser33/37/Thr41-β-catenin (p-Ser33-β-catenin), and non-p-Ser45-β-catenin (active β-catenin) were from Cell Signaling (Danvers, MA). Antibodies against p-Tyr1022-Jak1 and p-Tyr1007-Jak2 were purchased from Bioworld Technology (St. Louis Park, MN). Antibodies against Rock1, Rock2, Gsk3β, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-actin, and Lamin B were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Antibody against alkaline phosphatase (Alp), bone sialoprotein (Bsp), Lef1, Nestin, p-Ser1366-Rock2, and p-Tyr216-Gsk3β and recombinant human active Jak2 and Gsk3β were from Abcam (Cambridge, UK). The IRDye 680, 700, and 800 s antibodies were from LI-COR Bioscience (Lincoln, NE). Alexa555- and Alexa488-conjugated secondary antibodies were from Molecular Probes (Eugene, OR). Recombinant mouse active Wnt3a (rWnt3a), Wnt5a (rWnt5a), and Dkk1 (rDkk1) were from StemRD (Burlingame, CA). Y27632 and Fasudil were from Selleckchem (Huston, TX) and Pyridone 6 was from Medchem Express (Monmouth Junction, NJ).
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2

Affinity Purification and Immunoblotting Assay

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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. Rabbit polyclonal antibodies against LCMV NP were raised against NP protein (aa1-558).
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