Cell viability of M049 colon spheroids was performed by seeding the cells in 96-well plates (1000/ well) in stem cell medium in the presence of HGF (20 ng/ml). The following day, spheroids were treated with increasing concentrations of the fusion proteins (28,140 and 700 nM). After 4 days of treatment, cell number was determined using CellTiter-Glo (Promega Corp., Madison, WI, USA) with a VICTOR X4 multi-label plate reader.

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