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. 2012 Nov 14;304(4):F367–F375. doi: 10.1152/ajprenal.00537.2011

Fig. 3.

Fig. 3.

Af9 binds to +78/+92 of αENaC in vitro. Gel shift (autoradiograph to the left) in which a biotinylated duplex probe was incubated with 10 μg of mIMCD3 cell nuclear extracts alone (lane 2) or in the presence of a 40-fold molar excess of unlabeled wild-type (WT) cold probe (lane 3) or mutant cold probe (Mut; lane 4). Binding reactions were also performed in the absence of nuclear extracts as a control (lane 1). Antibody interference assays (autoradiograph to the right) were performed using increasing concentrations (0.2–2 μg) of rabbit polyclonal Af9 antibody or 2 μg IgG as control. Af9 antibody decreased formation of the DNA-protein complex, whereas IgG had no effect. Gels are representative of 4 experiments each for the gel shift and for the antibody interference assays.