Persistent binding of CDP/cut to CDP/cut site in the LF promoter.

(A) Electrophoretic mobility shift analysis was carried out using³²P-labeled double-stranded oligos encoding the CDP/cut site in the LF promoter (DM2 probe, lane 1). Addition of nuclear extracts prepared from uninduced (NB4, U, lane 2) and 48-hour ATRA-induced (NB4, 2d ATRA, lane 3) NB4 cells resulted in the formation of specific protein-DNA complexes (arrows), which were specifically competed away by the addition of a 100-fold molar excess of unlabeled self (lane 4, 100 × DM2), or known CDP/cut binding oligos (ie, NCAM; lane 5). (B) Binding of CDP/cut to the C/EBPε promoter. EMSA analysis was carried out using ³²P-labeled double-stranded oligos encoding the CDP/cut site in the C/EBPε promoter. Addition of nuclear extracts prepared from uninduced (NB4, U, lanes 2,5) and 48-hour ATRA-induced (NB4, 2d ATRA, lane 3) NB4 cells resulted in the formation of specific protein-DNA complex (arrow), which was specifically competed away by the addition of a 100-fold molar excess of unlabeled self (lane 4, × 100 DM2), or by the addition of increasing concentrations of a known CDP/cut binding site (ie, NCAM; lanes 6-8).