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FTC-133 and XTC-1 form tumor spheroids in suspension culture. To generate spheroids, FTC-133 and XTC-1 cells were plated in 24-well Ultra Low Cluster plates (Costar, Corning, NY) at a concentration of 4 × 104 cells per well and incubated at 37 C in 5% CO2 for 2 wk with media changed every 2–3 d. After 2 wk culture, tumor spheroids were photographed under the microscope and treated with Mercufenol. For each round of testing, quadruplicate wells were used for each testing concentration. The tumor spheroids were continuously treated for 2–3 wk and photographed under the microscope. For each compound and each cell line tested, the experiments were repeated at least three times