The human pancreatic cancer cell line PANC-1 obtained from the American Type Culture Collection, were maintained in Dulbecco's Modified Eagle Medium (DMEM) (Nissui, Tokyo, Japan) supplemented with 10% Fetal bovine serum (FBS) (biowest, Nusille, France), 2% L-glutamine, 1% antibiotics, and 1% MEN Non-Essential Amino Acids Solution (NEAA).
Lactate production in PANC-1 cell was measure of as follows; Cells were seeded at 8x104 cells/well to 24 well plate and culture 24 hours. The culture media were collected at 0, 3, and 6 hours and lactate determination used for lactate assay kit II. Oxygen consumption rate (OCR) was examined with a Seahorse Extracellular Flux Analyzer model 24XF (Seahorse Bio Science, MA). Cells were seeded at 4 x 104 cells/well in 24 well plates culture 24 hours before measurement. Effect of AG on mitochondrial respiration was examined by using XF Mito Stress Test Kit (Seahorse Bioscience, MA) according to the protocol recommended by the supplier.
Lactate production in PANC-1 cell was measure of as follows; Cells were seeded at 8x104 cells/well to 24 well plate and culture 24 hours. The culture media were collected at 0, 3, and 6 hours and lactate determination used for lactate assay kit II. Oxygen consumption rate (OCR) was examined with a Seahorse Extracellular Flux Analyzer model 24XF (Seahorse Bio Science, MA). Cells were seeded at 4 x 104 cells/well in 24 well plates culture 24 hours before measurement. Effect of AG on mitochondrial respiration was examined by using XF Mito Stress Test Kit (Seahorse Bioscience, MA) according to the protocol recommended by the supplier.