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HCT116 cells (5x103) were seeded in 200 μL of DMEM medium in 96-well U-bottom ultra-low attachment plates (Corning). The plates were incubated at 37°C in 5% CO2 for 5 days until the mean diameter of spheroids reached approximately 600 μm. Then, 100 μL of medium was replaced with fresh medium containing drugs and spheroid diameter was measured over 9 days. Spheroids were imaged and analyzed using GelCount Tumor Colony Counter (Oxford Optronix). At least 4 spheroids were measured per each treatment condition.
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This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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