Incucyte zoom kinetic live cell imaging

Cellular proliferation was assessed using the CellTiter 96 MTS cell Proliferation assay (Cat. #G109A, Promega, Madison, WI, USA). Samples were read at 490 nm in a Synergy 2 microplate reader (BioTek, Winooski, VT, USA). Cellular size, count and proliferation rates were also determined using the IncuCyte Zoom Kinetic Live Cell Imaging (Essen BioScience, Ann Arbor, MI, USA). IncuCyte cell recognition software calculated values based on cellular size over time, cellular count per field of view or percentage of cell confluence over time.

Cell proliferation was assessed by CellTiter 96 MTS Proliferation assay (Cat. # G109A, Promega) and IncuCyte Zoom Kinetic Live Cell Imaging (Essen BioScience). For MTS, samples were read at 490 nm in a Synergy 2 microplate reader (BioTek). Proliferation rates were determined using IncuCyte based on percentage of confluence over time. For migration, cells were plated in 96 well ImageLock Plates (Essen Bioscience). A scratch was made using a 96-pin WoundMaker (Essen Bioscience). For invasion, 96 well ImageLock Plates were coated overnight with 100 μg/mL Matrigel Basement Membrane Matrix (Cat. # 356231, Corning). The following day, Matrigel was aspirated and 25,000 cells were plated per well and allowed to adhere prior to making a scratch with the 96-pin WoundMaker. 50μL of Matrigel Matrix (8 mg/mL) was then added to the wells, and covered in 100 μL media containing the appropriate treatments (refer to figure legend for details). Migration and invasion were quantified using the “Relative Wound Density” metric generated by IncuCyte software.