Fig. 14 In vitro binding of TEF to the LRM D-box in an EMSA assay.
A 32P-labeled LRM probe was incubated with in vitro–synthesized TEF protein, in the presence or absence of specific unlabeled competitor DNAs. Reticulocyte lysate (RL) alone was used as a control for unspecific binding between RL proteins and the probe (Lane 1). Then TEF without competitor (Lane 2), TEF with the LRM as competitor (Lane 3), TEF with the LRM D-box as competitor (Lane 4), TEF with an Sp1 site as competitor (Lane 5), TEF with an LRM probe carrying a mutated D-box as competitor (Lane 6), and TEF with the LRM E-box as competitor (Lane 7). Black arrow indicates the TEF/LRM complex. An asterisk indicates the free LRM probe. These results indicate that TEF binds to the LRM-D-box.