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. Author manuscript; available in PMC: 2015 Jan 20.
Published in final edited form as: Virology. 2013 Dec 20;449:297–303. doi: 10.1016/j.virol.2013.11.031

Figure 7.

Figure 7

Identification of nucleotides in NSBEs 1 and 2 critical for NS1N binding. (A) Lanes 1-8, NS1N incubated with the 28-bp NSBE1+2-1AA, NSBE-1+2-2TT, NSBE1+2-3TT, NSBE1+2-4AA, NSBE1+2-5TT, NSBE1+2-6TT, NSBE1+2-7AA and NSBE1+2-8AA DNA, respectively (for nomenclature of DNA molecules, refer to Table 1). Lane 9, the 28-bp NSBE1+2+3 wt DNA alone. Lane 10, NS1N incubated with the 28-bp NSBE1+2+3 wt DNA. The potential nucleoprotein complexes are indicated with an arrow. Lane M, DNA marker. (B) A plot showing effects of point mutations in NSBEs on NS1N binding, which combines EMSA results for the eight DNA molecules in (A) (see Table 1 for actual sequences of DNA molecules used in the assay). The vertical axis is semi-quantified binding strength based on EMSA results. The horizontal axis shows the nucleotide sequence of the region of NSBEs 1 to 3 (upper row in black) as well as the nucleotides they were mutated to (bottom row in red). The effect of mutation in NSBE3 was based on a single EMSA result using the 28-bp NSBE3-1to8 DNA, in which all the eight nucleotides were mutated (Table 1; Figure 4, lane 6).