Figure 5
In vitro Glut1 deletion induces pro-apoptotic Bim expression and sensitizes B-ALL to cell death stimulus. (a and b) WT Cre-ER and Glut1fl/fl Cre-ER B-ALL cells treated with vehicle or 4-OHT for 96 h followed by 48 h of culture without 4-OHT and (a) examined by immunoblot on day 6 or (b) analyzed by flow cytometry for survival over time. (c) WT Cre-ER and Glut1fl/fl Cre-ER B-ALL cells were cultured with vehicle or 4-OHT for 96 h, washed, then cultured an additional 48 h alone or with addition of Dasatinib (50 nM), and cell viability was determined over time by flow cytometry. (d) Apoptosis in Glut1-deleted cells with or without Dasatinib treatment was assessed by annexin V/PI staining. Cells were treated with vehicle or 4-OHT for 96 h and apoptosis was assessed by annexin V/PI staining (left panel). After 96 h of culture with vehicle or 4-OHT, cells were washed and cultured for an additional 48 h alone or with Dasatinib (50 nM). Cell apoptosis was assessed at the end of the 48 h (right panel). Ten μM pan caspases inhibitor Q-vd-oph was added in some cell cultures as indicated. Gray bar, annexin V+/PI− cell percentage. Black bar, annexin V+/PI+ cell percentage. Means and S.D. are shown for triplicate samples from representative experiments repeated three or more times. *P<0.05, **P<0.005, ***P<0.001, ****P<0.0001
