Gel mobility shift analyses using nuclear extracts from HeLa cells and NIH3T3 cells with WT h-TR
1 on the (A) Lys- and (B) ME-TREs
An equal amount of in vitro translated receptor was incubated with labeled Lys-TRE and 5.0 µg of HeLa or 3T3 nuclear extract and electrophoresed. Exposure of the dried gels was kept constant between experiments at 14 hr. The 3T3 section in Panel A was exposed for 2.5 days in order to bring out the heterodimer and homodimer bands. Note that a nonspecific band (NS) runs just above the homodimer band in the panel with HeLa nuclear extract and is not competed away by specific competitor. (▷) The lower band in Lane 5 likely represents a faster migrating heterodimer band competed away by specific competitor (Lane 6), and overlaps a nonspecific band. Note that in Panels A and B, all lanes represent each of two separate gels, respectively.