Analysis of PLZF protein expression in the inducible clones. (a) Western blot prepared from whole-cell extracts of human Jurkat cells stably transfected with pJEF4 (clone vector 4) or pJEF4-PLZF (clones PLZF 3 and 15), obtained 72 h after washing the cells free of dox and reculturing in the presence (lanes 1, 3, and 5) or absence (lanes 2, 4, and 6) of dox, incubated with a PLZF mAb. A band at the expected molecular mass of 80 kDa was detected in clones 3 and 15 after dox withdrawal. (b) Flow cytometry analysis of the clones by using the PLZF mAb and secondary labeling with anti-mouse IgG coupled to phycoerythrin. (c) Western blot of whole-cell extracts of clone PLZF 15 showing the kinetics of induction of PLZF expression after dox withdrawal.