Figure 2. FK866 produces metabolic collapse in pancreatic cancer cells.

A, Panc-1 cells were kept in 1% FBS for 48 hours and then treated with FK866 (10 nM) or vehicle. 48 hours later media was changed to glucose-free DMEM supplemented with 5 mM D-[C¹³] glucose. Glucose consumption and glycolytic intermediates were determined using NMR spectroscopy, and glycolytic intermediates determined. B, cells were kept in 1% FBS for 48 hours and then treated with FK866 (10 nM) or vehicle for 48 hours. The medium was replaced by RPMI phenol red and serum-free and aliquots of cultured medium were collected for lactate release evaluation. C, O₂ consumption rates were measured by high resolution respirometry. After 48 hours treatment with 50 nM of FK866 or vehicle, the cells resuspended in serum-free DMEM, and ETS-stimulated respiration (maximum respiration) was measured. Results represent the mean ± SD of three independent experiments. D, Cells were kept in 1% FBS for 48 hours then treated with FK866 or vehicle for 48 hours before ATP measurements. Values are means ± SD of three independent experiments. Cell lysates of PaTu8988t cells treated with 10nM FK866 for 24 hours were immunoblotted with anti-p-AMPK and AMPK antibodies. * indicates p<0.05