WAVE3 Knockout in Breast Cancer Cells

BC cell lines were obtained from American Type Culture Collection and maintained according to the manufacturer’s protocols. Cell lines were also routinely authenticated by short tandem repeat DNA fingerprinting analysis. WAVE3-KO cells were generated by lentiviral transduction as described previously^{34 (link)}. We used two different and verified WAVE3-specific single guide RNAs (sgRNAs) for each of the human and mouse BC cell lines and a scrambled (SCRAM) sgRNA^{34 (link)}. We used the following reagents: platelet-derived growth factor (PDGF; Millipore) (100 ng/ml for 10 min); TFG-β (R&D; 5 ng/ml for 20 min); EGF (Gibco) (100 ng/ml for 10 min); LY294002, ZD1839, SB431542, LY2109761, and Alpelisib (Selleckchem); AG1296 (Apexbio Technology); AKT1/2 Kinase-inhibitor (Sigma-Cat.# A6730; 10 µM for 4 h); and Matrigel (Corning; 4.5 mg/ml).