XB-IMG-135056
Xenbase Image ID: 135056
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Fig. 2. GATA-4 binds specifically to a GATA cis-element. Gel
mobility-shift assays were performed using a labeled oligomer
probe (P) containing a GATA cis-element from the chicken aDglobin
promoter [TE72/73 (Evans and Felsenfeld, 1991)]. The
probe was incubated in the absence of added extract (-) or with 1
or 2 ml (left to right) of nuclear extract prepared from chicken
erythroid cells containing cGATA-1 (RBC), COS cells transfected
with the pXM expression vector without an insert (COS), or COS
cells transfected with the xGATA-4 expression plasmid (xG4). All
other samples included 2 ml of the xG4 nuclear extract in addition
to competitor. Competitors were a 30- or 60-fold excess (left to
right) of unlabeled TE72/73 oligomer as a specific DNA
competitor (Sp), the same amounts of unlabeled TE78/79 (Evans
and Felsenfeld, 1991), which is identical to TE72/73 except for a
mutation of the GATA motif, as a non-specific DNA competitor
(Ns), 1 or 2 ml of pre-immunized rabbit serum (pb), 1 or 2 ml of
polyclonal rabbit anti-GATA-4 serum (pAb), 1, 2 or 4 ml of
protein A affinity-purified monoclonal antibody directed against
GATA-4 (mAb). Note that the complex formed by xGATA-4
(arrow) is of slightly higher mobility compared to that formed
with cGATA-1 (arrowhead), consistent with the larger molecular
weight of xGATA-4. The asterisk indicates a complex formed to a
partial degradation product of GATA-4, which retains binding
activity. Image published in: Kelley C et al. (1993) Copyright © 1993. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View |