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. 2016 Nov 28;28(12):2916–2936. doi: 10.1105/tpc.16.00609

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Figure 5. — NKD1 and NKD2 DNA Binding.

(A) and (B) SAAB-MEME derived BCSs for NKD1ID (A) and NKD2ID (B).

(C) Oligonucleotide sequences used as probes for EMSAs. The wild type contains the consensus binding sequence, while M1 to M8 contain base substitutions indicated by magenta lettering. Binding, relative to the wild type for a given protein is indicated by +++ (>66%), ++ (33 to 66%), + (<33%), and – (no detected shift).

(D) to (G) EMSAs.

(D) and (F) EMSA of consensus (wild type) and mutant probes for NKD1-ID (D) and NKD2-ID (F) proteins.

(E) and (G) Competition assays using labeled wild-type probe, and 50-, 100-, or 500-fold excess of unlabeled wild type, M2, or M8 oligonucleotide, incubated with NKD1ID (E) or NKD2ID (G) protein. The + indicates reaction with no competitor, and – indicates negative control with GST protein instead of NKD. Overexposed images, additional EMSAs, and negative controls using purified GST are shown in Supplemental Figures 8 and 9.