Anti g3bp1

The interaction between G3BP1 and β-catenin proteins was assessed using a Co-IP assay. SW620 and RKO cells infected with Vector-G3BP1 or Vector-negative control (NC) were rinsed with cold PBS and lysed in IP lysis buffer, followed by centrifugation at 10,000 × g at 4°C for 30 min. Then, 200 µg proteins from each sample were incubated with Dynabeads^® protein G (Thermo Fisher Scientific, Lnc.) for 1 h at room temprature, and incubated with 2 µg of anti-G3BP1 (cat. no. ab181150; Abcam) or anti-β-catenin (cat. no. ab16051; Abcam) antibody overnight at 4°C. Anti-IgG antibody (cat. no. ab182931) was used as a negative control. This was followed by incubation with Dynabeads^® protein G for another 1 h at room temperature. Subsequently, the immunocomplex was washed five times with IP lysis buffer (Thermo Fisher Scientific, Inc.) and then subjected to western blot analysis with anti-β-catenin (cat. no. MA1-300; Thermo Fisher Scientific, Inc.) or anti-G3BP1 (cat. no. ab56574; Abcam) antibodies at 1:2,000 dilution.