Targeting glutaminase activity inhibits AML cell proliferation. (A) OCI-AML2 and MOLM-14 shGLS1#5 leukemic cells were seeded at 5 × 105 cells/mL with or without doxycycline; viable cells were counted manually following trypan blue staining on days 3, 5, and 7. Each experiment was performed independently ≥3 times. (B) OCI-AML2, MV4-11, MOLM-14, HL-60, and OCI-AML3 cells transfected with shGLS1#5 or #7 were seeded at 5 × 105 cells/mL with or without doxycycline; viable cells were counted following trypan blue staining on days 3, 5, and 7. Each experiment was performed independently ≥3 times. (C) OCI-AML2 and MOLM-14 shGLS1#5 leukemic cells were seeded at 5 × 105 cells/mL with or without doxycycline, and αKG (5 mM) and viable cells were counted following trypan blue staining on day 7. (D) A total of 10 cell lines were seeded at 5 × 105 cells/mL with or without CB-839 (1 µM); viable cells were counted following trypan blue staining on days 3, 5, and 7. (E) OCI-AML2 cells were seeded at 5 × 105 cells/mL with or without CB-839 (1 µM) and αKG (5 mM); viable cells were counted following trypan blue staining on days 3, 5, and 7. (F) CD34+ HPCs from 4 healthy donors were seeded at 5 × 105 cells/mL with and without CB-839 (1 µM); viable cells were counted following trypan blue staining on days 2, 3, 4, and 5. *P < .05, **P < .01, ***P < .001.
