4.10. Immunoprecipitation with Synchronized U2OS Cells

Two point five times ten to the six U2OS cells were seeded on T175 flasks two days before dexamethasone shock. Cell synchronization was performed with 100 nM dexamethasone (D4902, Sigma) for 30 min at 37 °C. Cells were collected after 26 h, 30 h, 34 h, 38 h, 42 h, and 46 h after dexamethasone treatment. Immunoprecipitations were performed from 4 mg whole cell protein extract with 4 µg CSNK1D (ab103955, Abcam) or 4 µg GAPVD1 antibody (NBP1-19156, Novus Biologicals).