TNBC Cell Line Generation and Characterization

TNBC Cell lines were acquired from the American Type Culture Collection (ATCC; Manassas, VA), and maintained according to the manufacturer’s established protocols. Cell lines were also routinely authenticated by short tandem repeat DNA fingerprinting analysis. WAVE3-deficient (W3-KO) cells were generated using two different and verified sgRNAs targeting the human WAVE3 and the mouse Wave3 genes by lentiviral transduction as described previously [11 (link)]. A scrambled (SCRAM) sgRNA was used as negative control [11 (link)]. Generation of the WAVE3-KO cells overexpressing wildtype (WT) or phosphomutant (Y4) were described previously [17 (link)]. GFP-tagged WAVE3 constructs were generated as described previously [17 (link)]. The GFP-recombinant vector or the empty GFP expression control vector was used for stable transfections using standard protocols. We used the following reagents: Doxorubicin, docetaxel and cisplatin, purchased from Sigma (St. Louis, MO) as powders, were dissolved in DMSO and used as described previously [12 (link)]. For proteasome inhibition, cells were treated with GM6001 (Sigma) at 20 µM for 24 h. Gel electrophoresis reagents were from Bio-Rad (Hercules, CA).