Compound used as Wnt signal pathway activator, and preparation and application thereof
A technology of compounds and signals, applied in medical preparations containing active ingredients, anti-inflammatory agents, drug combinations, etc., can solve the problem that there is no obvious progress in research.
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[0082] 1. Experimental materials
[0083] 1) Plasmid
[0084] TOPFlash plasmid was purchased from Millipore Corporation.
[0085] GFP plasmid was purchased from Millipore Corporation.
[0086] 2) Antibody
[0087] The antibody of β-actin (sc-47778) was purchased from Santa Cruz; the antibody of SP1 (S9809) was purchased from Sigma; the antibody of β-catenin (610154) was purchased from BD Biosci.Pharmingen.
[0088] 3) Reagents
[0089] Protease inhibitors (complete protease inhibitor mixed tablet) and NP-40 were purchased from Roche; Lipofectamine and PLUS transfection reagent were purchased from Invitrogen;
[0090] Wnt3a conditioned medium was prepared from Wnt3a cell line (CRL-2647, purchased from ATCC cell bank in the United States): Wnt3a cells were cultured in DMEM cell medium supplemented with 10% calf serum for 4 days, and then the supernatant medium was collected by centrifugation.
[0091] The control conditioned medium was prepared from the L cell line (CRL-2648,...
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