MCF10A cells (American Type Culture Collection) were cultured in Dulbecco’s modified Eagle’s medium (DMEM)/F12 media (Thermo Fisher Scientific) supplemented with 5% horse serum (Thermo Fisher Scientific), epidermal growth factor (EGF; 20 ng/ml; PeproTech), insulin (10 μg/ml; Sigma), hydrocortisone (500 ng/ml), and cholera toxin (100 ng/ml; Sigma). MCF10A NLS copGFP and MCF10A paxillin-mCherry stable lines were prepared by lentiviral transduction using NLS copGFP pCDH and paxillin-mCherry pLV hygro tetOn plasmids, respectively. MCF10A F-tractin–EGFP (enhanced green fluorescent protein) stable lines were prepared using a transposon-based method with an F-tractin–EGFP plasmid. The backbone pPB puro tetOn was modified by swapping a eukaryotic antibiotic resistance marker from puromycin to zeocin. Then, the F-tractin–EGFP plasmid was prepared by inserting F-tractin–EGFP from F-tractin–EGFP C1 into pPB tetOn using Bam HI and Eco RI restriction sites. National Institutes of Health (NIH) 3T3 cells and GM00637 (Cornell Cell Repository) cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific). PBMCs were cultured in RPMI (Thermo Fisher Scientific) supplemented with 1% GlutaMAX (Thermo Fisher Scientific) and 10% FBS (Thermo Fisher Scientific).

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