Cells were plated at equal concentrations with varying concentrations of the inhibitor and in triplicate wells. Cells were counted, the concentration was calculated every second/third day, and the cells were replated at even cell concentrations. A growth curve was plotted for cumulative growth according to the dilution at each time point. Doubling time was calculated from fitted semi-logarithmic slope (Nonlin). Fit using least squares in GraphPad Prism software by log2/slope and Student’s two-tailed t test was used to test for significance. For cell cycle assays (Lp30 cells), cells were plated with 30 μM inhibitor or DMSO for 24 hours. Cells were harvested, and an equal number of cells were fixed in 2% paraformaldehyde, permeabilized in 0.1% saponin, stained with 4′,6-diamidino-2-phenylindole (DAPI), and analyzed by flow cytometry (LSR-II, BD Biosciences). For apoptosis assay (Lp30 cells), cells were plated with 30 μM inhibitor or DMSO for 72 hours, harvested, washed and stained using a PE-annexin V kit (BD Biosciences), and analyzed by flow cytometry (Accuri, BD Biosciences). All flow cytometry analyses were run with FlowJo software V.10.

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