Cell Viability Assay with Inhibitors

Cell growth over time was measured using the PrestoBlue Cell Viability Reagent (Invitrogen). Briefly, 67NR, 5637, SW480 or MDA-468 cells were seeded in 96-well plates (2,500 cells/well) and exposed to ATX-101 (25 amino acid cell-penetrating APIM-containing peptide [21 (link)], Ac-MD-RWLVK-W-KKKRK-I-RRRRRRRRRRR, Invitrogen, Sweden) (4-12 μM), ATX-A (Ac-MD-RALVK-W-KKKRK-I-RRRRRRRRRRR, Invitrogen, Sweden) (6 μM), AEE788 (EGFR/HER2/VEGFR inhibitor, Selleck Chem) (0.5-1 μM), Bafilomycin A1 (10 nM) Cell Signaling, #54645, or cMet inhibitor (PHA-665752, Sigma, 2 μM) until harvest at day three. Data displayed are percentage viability relative to untreated control in one representative experiment out of three biological replicas demonstrating the same trend.