Regulation of β-Catenin Acetylation

SR3029, D4476, MG149, NU9056, MG132 and cycloheximide (CHX) were purchased from MedChemExpress (MCE, Monmouth Junction, NJ, USA). The following primary antibodies were used: anti-CK1δ, anti-β-catenin, anti-p300, anti-acetyl-lysine, IgG (Santa Cruz Biotechnology, Heidelberg, Germany), anti-CBP, anti-acetyl-β-catenin (K49), anti-phospho-β-catenin (S45), anti-Flag, anti-V5, anti-CK1ϵ, anti-GFP, anti-mouse IgG (Cell Signaling Technology, Danvers, MA), anti-Tip60 (Abcam, Cambridge, MA, USA), and anti-GAPDH antibody (Proteintech, Chicago, IL, USA). The Tip60-Flag, PCAF-Flag, p300-Flag and pEGFP-N1 plasmids were purchased from Vigenebio (Weizhen, Shandong, China). The SuperTopFlash reporter plasmid was provided by Karl Willert (University of California at San Diego, La Jolla, CA, USA). The expression plasmids encoding β-catenin, pCMXβgal (β galactosidase, β-gal), CK1α-V5, CK1δ-V5, CK1ϵ-V5, CK1α-Flag, CK1δ-Flag, CK1ϵ-Flag, CK1γ-Flag have been described previously (35 (link), 36 (link)). For the construction of GFP-tagged CK1δ, the cDNAs encoding human CK1δ was amplified by PCR and subcloned into the BamHI/EcoRI site of pEGFP-N1 vector using ClonExpress Ultra One Step Cloning Kit (Vazyme, Nanjing, China). The primer sequences used are as follows: CK1δ-GFP-sense, 5’-CTCGAGCTCAAGCTTCATGGAGCTGAGAGTCGGGAAC-3’; CK1δ-GFP-antisense, 5’-CTCACCATAAGGTGGCGACCGGTGTAGGTGCGTCGTG.