Guanylate cyclase receptor agonists for the treatment of tissue inflammation and carcinogenesis
A technology of guanylate cyclase and receptor antagonists, which is applied in the field of guanylate cyclase receptor antagonists, and can solve the problems of decreased production of uroguanylin and guanylin
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[0140] Materials and Methods
[0141] Cell culture: Human T84 colon tumor cells were obtained from the American Type Culture Collection (ATCC) at passage 52. Cells were grown in a 1:1 mixture of Ham's F-12 medium and Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum, 100 units per milliliter of penicillin, and 100 micrograms per milliliter of streptomycin. Replace with fresh medium every two days, and split the bottles when the confluence rate is about 80%.
[0142] T84 Cell Based Assay for Determining Intracellular cGMP Levels: Phthaloids were custom synthesized by Multiple Peptide Systems, San Diego, CA and Princeton Biomolecules Langhorne, PA. The biological activity of the synthetic peptides was determined according to previous reports (15). The brief steps are as follows. The confluent monolayer cells in the 24-well culture plate were washed twice with 250 microliters of DMEM containing 50 mM HEPES (pH7.4), and washed with 250 microlit...
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