Figure 5

The transcription factor Elf-1 binds to the -360 Ets site in the LAT promoter. A. A DNA probe encompassing the Ets site centered at -360 (-360 Ets wt) was labeled with ³²P and incubated with nuclear extracts from Jurkat T cells in the absence or presence of 100-fold molar excess of competitor DNA. Samples were subsequently loaded onto a nondenaturing polyacrylamide gel, electrophoresed, and then subjected to autoradiography. For competition experiments, the unlabeled competitor DNA was the same sequence as that of the probe (self wt), the same sequence as the probe except containing point mutations within the Ets site (self mutEts), the consensus sequence for Ets family members (cons. wt) or the consensus sequence except for point mutations within the Ets site (cons. mutEts). The arrow indicates the specifically bound complex. B. Experiments were conducted as in Fig. 5A except that the indicated antibody was added in place of competitor DNA. C. A longer exposure of the lane containing the Elf-1 antibody. The supershifted complexes are indicated with arrows.