Figure 5

MYC-dependent osteogenic sarcoma cells are dependent on aminotransferases. Doxycycline (20 ng/mL) was added to MYC-dependent osteogenic sarcoma cells and incubated for 48 hours. (a) Cells were incubated without glutamine for one hour. OCR was measured and subsequently after the addition of glutamine (4 mM) followed by AOA (1 mM) and DMK (5 mM). N = 7 ± SEM * P < 0.05 compared to no glutamine. (b) Protein expression of GLS1, GPT2, and GOT2 in mitochondrial and cytoplasmic fractions in cells treated with doxycycline. VDAC1 and tubulin are loading controls for mitochondrial and cytosolic fractions, respectively. (c) Cell proliferation was assessed after 48 hours treatment with AOA (1 mM). N = 3 ± SEM * P < 0.05 compared to 0 mM DMK with 1 mM AOA. (d) Cell death assessed after 48 hours of treatment with AOA (1 mM) in the presence of DMK (0–5 mM). N = 6 (control condition) and N = 3 (for experimental conditions) ± SEM * P < 0.05 compared to 0 mM AOA.