Colon Cancer Cell Treatment Protocol

SW48 colon cancer cells (ATCC – CCL-231) were maintained in RPMI-1640 (Gibco) supplemented with 10% fetal bovine serum (FBS) (#TMS-013-B, Millipore) and 1% penicillin-streptomycin (#15070, Gibco). Cells were grown at 37°C in 5% ${\mathrm{CO}}_2$ and passaged at 70% confluency. For all experiments, a treatment of 250-nM ABT-263 (ApexBio cat# A3007), 10-nM TAK-228 (LC Laboratories cat.# I-3344), or the combination was used. These doses are based on physiologically relevant blood serum levels of ABT-263³⁷ (link)^,38 (link) and on a WST-1 dose response curve for TAK-228, where 10 nM was shown to decrease proliferation of cells by $>50\%$ compared to control.³⁹ (link) Cells were exposed to treatment for 24 h.