Fig. 1. ETV7 induces rapamycin resistance in mouse WT and Arf^−/− pre-B cells.

(A) Cell lysates from WT and Arf^−/−mouse pre-B cells (ARF^−/−) expressing GFP (Vec) or ETV7 (ETV7) immunoblotted for mTOR, p-P70S6K^Thr389, total P70S6K, p–4E-BP1^Thr37/46, p–4E-BP1^Ser65, p–4E-BP1^Thr70, total 4E-BP1, total 4E-BP2, p-PDK1^Ser241, total PDK1, p-AKT^Thr308, p-AKT^Ser473, total AKT, p-NDRG1^Thr346, total NDRG1, total eIF4E, ETV7, and tubulin as a loading control. (B) Proliferating Arf^−/− mouse pre-B cells transduced with murine stem cell virus (MSCV)–internal ribosomal entry site (IRES)–green fluorescent protein (GFP) (vector) or MSCV-ETV7-IRES-GFP (ETV7) were treated with increasing amounts (0.1, 0.3, 1.3, 10, 30, 100, 300, and 1000 ng/ml) of rapamycin or AZD-8055 for three population doublings. Cell densities (percent control) were plotted as the percentage of cells treated with vehicle. Data are means ± SEM from three independent experiments. (C) Cell lysates of proliferating Arf^−/− mouse pre-B cells transduced with MSCV-IRES-GFP (vector) or MSCV-ETV7-IRES-GFP (ETV7) were immunoblotted after treatment of the cells with increasing amounts (0.1, 0.3, 1.3, 10, 30, 100, 300, and 1000 ng/ml) of rapamycin for three population doublings. The blots were probed for total mTOR, mTOR^Ser2448, p-P70S6K^Thr389, total P70S6K, p–4E-BP1^Thr37/46, total 4E-BP1, total 4E-BP2, p-GRB10^Ser501/503, total GRB-10, p-NDRG1^Thr346, total NDRG1, p-AKT^Thr308, p-AKT^Ser473, total AKT, p-ERK^{Thr202/Tyr204}, and total eIF4E.