Anti ctnnb1

WB was performed as previously described [9 (link)]. After incubation with the secondary antibody, the proteins were detected by chemiluminescence (Millipore Germany). Primary antibodies used in this study were anti-ALX4 (1:1000; Santa Cruz Biotechnology), anti-CTNNB1 (1:700; Santa Cruz Biotechnology), anti-p-CTNNB1 (1800; Santa Cruz Biotechnology), anti-GSK3β (1:800; Santa Cruz Biotechnology), anti-c-Myc (1:1000; Santa Cruz Biotechnology), anti-CCND1 (1:1000; Santa Cruz Biotechnology), anti-MMP7 (1:1000; Abcam) and anti-GAPDH (1:2000; Beyotime China). The siRNA targeting human GSK3β was purchased from Riobio (stQ0004712–1).

Cells were lysed using lysis buffer (50 mM HEPES, 5 mM EDTA, 50 mM NaCl, 1% Triton X100, 50 mM NaF, 10 mM Na2P4O7, 1 mM Na3VO4, 5 μg/mL aprotinin, 5 μg/mL leupeptin, 1 mM PMSF, and a protease inhibitor cocktail). Lysates containing equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene difluoride membrane (Bio–Rad, Hercules, CA, USA). The membrane was blocked with a 5% skim milk solution and incubated with the following antibodies: anti-ADAR1, anti-CTNNB1, anti-GAPDH, anti-MSI2, anti-MET, anti-SLC38A4 (Santa Cruz Biotechnology, Dallas, TX, USA), and anti-FLAG-Tag (Cell Signaling Technology, Danvers, MA, USA). An Immobilon™ Western blot detection system (Millipore) was used to detect bound antibodies. The intensities of the western blot bands were quantified using LAS-4000 (Fuji Photo Film Co., Tokyo, Japan).