Fig 1. Effect of ethyl pyruvate and ethyl lactate on T. brucei cells proliferation.

A cell proliferation/viability assay was conducted in 96-well cell culture plates containing 2x104 cells, medium and ethyl pyruvate in a volume of 200 μl. After 24 hrs of incubation the AlamarBlue cell proliferation reagent (20 μl) was added and the absorbance was read: (A) Ethyl pyruvate; (B) Ethyl lactate; (C) For the time-dependency test 2x105 cells/ml were cultured in 24-well plates in the presence of different concentrations of ethyl pyruvate at 0 mM (■), 1 mM (▲), 2.5 mM (x) and 5 mM (□). After certain time intervals aliquots were removed and the number of vital cells was counted. (D) For cell-recovery test 2x105 cells/ml were cultured in the presence of ethyl pyruvate at 0 mM (■), 1 mM (▲), 2.5 mM (▼), 5 mM (□) for 3 hrs in 24-well plates. Then, the medium was removed and the cells were replenished with fresh medium without the ethyl pyruvate and further cultured for 24 hrs. At the indicated time points aliquots were removed and vital cells were counted. Cells were cultured at 37°C containing 5% CO2 in a 100% humidified environment, * p<0.05.