Generation of Bcr-Abl and STAT5 inducible cell lines.
(A) Ton.B.210, Ton.B.STAT5, and Ton.B.1*6 cells were cultured in RPMI 1640 medium supplemented with 10% FCS and incubated 24 hours, with (+) or without (−) doxycyline. Cells were harvested and lysed, and the lysates were separated by SDS-PAGE. After transfer to a PVDF filter, the inducible expression was evaluated by immunoblotting with the specified antibodies. (B) 1 × 107 Ton.B.210, Ton.B.STAT5, or Ton.B.STAT51*6 cells were cotransfected with GAS-luc and pCMV-βGal constructs (25 μg each). Transfected cells were then split and incubated in the same medium with or without doxycycline for 24 hours. Cells were harvested and lysed for reporter gene assays as described in “Material and methods.” Results are reported as the activity in induced cells compared with the activity in noninduced cells for each cell line (% of control).
