Sh nc

To overexpress G3BP1, colon cancer cells were infected with the lentivirus vector (Vector-G3BP1; Shanghai GenePharma Co., Ltd.) using 5 µg/ml polybrene (Hanbio Biotechnology Co., Ltd.) with an MOI of 10, and the infected cells were incubated with G401 (100 µg/ml) for 14 days at 37°C to establish the stably infected cells.
To silence β-catenin expression, colon cancer cells were infected with the lentivirus vector [short hairpin RNA (sh)-β-catenin; OriGene Technologies, Inc.] and puromycin (100 µg/ml) was applied to select the stably infected cells at 37°C for 14 days.
To knockdown G3BP1 expression, colon cancer cells were transfected with the small interfering (si)RNAs targeting G3BP1 (si-G3BP1; OriGene Technologies, Inc.) using Lipofectamine^® 2000 reagent (Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. The sequences were as follows: i) si-G3BP1-1, 5′-CCACACCAAGATTCGCCAT-3′; ii) si-G3BP1-2, 5′-GGAGATTCATGCAAACGTT-3′; iii) si-G3BP1-3: 5′-GGAGGAGTCTGAAGAAGAA-3′; and iv) si-NC: 5′-CCAAACCTTAGCGCACCAT-3′. Vector-NC, sh-NC and si-NC (OriGene Technologies, Inc.) were used as the negative controls for Vector-G3BP1, sh-β-catenin and si-G3BP1, respectively. After 48 h of transfection/infection, the cells were collected for subsequent experiments.