Figure 3

The rate of glycolysis and dependency on glucose are increased in BRAT1 knockdown cells. (A) Culture media were harvested from control or BRAT1 knockdown cell cultures at indicated day and then glucose consumption (left) and lactate accumulation (right) were analyzed. (B) Control and BRAT1 knockdown cells were cultured with (glucose +) or without glucose (glucose -). Cells were harvested at indicated day and stained with PI without fixation. PI positive cells were detected as apoptotic/necrotic cells. Data were expressed as mean fluorescence intensity (ΔMFI = MFI of PI stained cells – MFI of unstained cells). (C) Cells were treated with or without 2DG (5 mM) for 24 h and then cellular apoptosis was detected with AnnexinV/PI double staining by flow cytometry. Flow cytometry data were representatives of two different experiments. (D) Culture media were daily changed and the number of living cells was counted after trypton blue staining.