Cells (10,000 cells per well) were seeded on uncoated bacteriological 35-mm dish (Coning Inc., Coning, NY, USA) with 3D Tumorsphere Medium XF (Sigma). Cells were cultured with or without PTE (10, 50, 100 μM). After 7 days, sphere images were captured on a computer and the sphere size was measured using NIH ImageJ software (version 1.52, NIH, Bethesda, MD, USA). We counted the number of all spheres in the culture well. The sphere size designated was a mean of all spheres in the well.
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