5 ethynyl 2 deoxyuridine edu incorporation assay kit

The primary antibodies and their working concentrations used in this study are listed in Supplementary Material Table 1.2.3. The Mitochondria Fractionation Kit (Cat# C3601) was purchased from Beyotime (Biotechnology, China). PDE2 Activity Assay Kit (Cat# ab139460), PKA Kinase Activity Assay Kit (Cat# ab139435), cAMP Assay (Cat# ab65355) and ODQ (Cat# ab120022) were all purchased from Abcam (Abcam, United Kingdom). Go 6983 (Cas# 133053-19-7) inhibitor was from MCE (MCE, USA). The CellTiter 96^® Aqueous One Solution Cell Proliferation Assay was purchased from Promega (Cat# G3581, Promega, USA) and 5-ethynyl-2’-deoxyuridine (EdU)-incorporation assay kit was purchased from Ribobio (Cat# C10310, Ribobio, China). Bay 60-7550 (Cat# HY-14992) and H89 (Cat# HY-15979) were purchased from Med Chemexpress (Med Chemexpress, USA).

The MTT assay was conducted to evaluate the cell growth rate. Cells were seeded in a 96-well plate (4000 cells/well) and incubated for 24 h with indicated concentrations of drug. The absorbance of each well was detected by a spectrophotometry at 570 nm wavelength. As for colony formation assay, CRC cells were seeded in a 24-well plate (500 cells/well) for individual treatments. After treatment of one week, 4% paraformaldehyde (PFA) was used to fix cells and crystal violet was used to dye the cells. After washing three times with water, dyed cells were dissolved in 0.1% Sodium dodecyl sulfate (SDS). Then, the absorbance was measured at 570 nm reference wavelength with microplate reader. The 5-Ethynyl-2′-deoxyuridine (EdU) incorporation assay kit (Ribobio) was used to detect cell proliferation as described previously 8 (link).