Fig. 5.

Effect of BC-LI-0186 on cancer cell death. a SW620 cells were treated with DMSO (Con), rapamycin (100 nM), BC-LI-0186 (10 μM), or 5-FU (10 μM) for 24 h. After cells were stained with PI and Annexin V, cells were separated and counted by FACS. b SW620 cells stably expressing RFP were treated with DMSO or 1.85 µM BC-LI-0186 in the presence of CellTox green. After 0, 24, 48 h, red (cell growth) and green (cell death) fluorescence images were acquired. c SW620 cells stably expressing RFP were treated with various concentration of BC-LI-0186 in the presence of CellTox green to monitor cell growth and death, simultaneously. The GI₅₀ (red line) and EC₅₀ (blue line) of BC-LI-0186 were determined by analyzing dose-response curves using GraphPad Prism tools. The insets represent the GI₅₀ and EC₅₀ values of BC-LI-0186. d SW620 cells were treated with DMSO (Con), BC-LI-0186 (10 μM), rapamycin (100 nM), or 5-FU (10 μM) for 24 h. Cells were analyzed by immunoblotting with the indicated antibodies. e SW620 cells were treated with 10 μM BC-LI-0186 for 0, 3, 6, 9, 12 h and analyzed by immunoblotting with the indicated antibodies. f SW620 cells were treated with DMSO (Con), BC-LI-0186 (0.1, 1, 10, 20 μM), rapamycin (100 nM), or 5-FU (10 μM). After 24 h, apoptosis was measured using Cellplayer caspase-3/7 reagent in the absence or presence of Q-VAD-FMK, which is an apoptosis inhibitor. g SW620 cells were treated with DMSO (Con), BC-LI-0186 (10 μM), rapamycin (100 nM), or 5-FU (10 μM) in the presence of CellTox green to measure cell death. After 24 h, green fluorescence was monitored in the presence of Q-VAD-FMK. The error bars represent mean ± S.D. (n = 3)