Integrin β5 Knockdown via Lentiviral Vectors

The methods for vector construction and generation of stable cell lines were described in our previous study [30 (link)]. The oligonucleotides to suppress integrin β₅ expression were designed by RiboBio (Guangzhou, China). After we verified their knockdown efficiency, they were cloned into lentiviral expression vector pLKO.1‐Pur (Addgene, Cambridge, MA, USA). The plasmids were verified by sequencing. Empty vector pLKO.1‐Pur carrying a scrambled shRNA served as a control. 293T cells were incubated with the constructed vectors, pMD2G and psPAX2 (Addgene), according to the manufacturer's protocol. 0.22 μm PVDF filters were used to filter 293T cells supernatant, and then, the supernatant was added into the plate to infect DLD1, HCT116, and HCT8 cells. The oligonucleotide sequences for vector construction are listed in Table S4.