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. 2015 Jan 27;167(3):991–1003. doi: 10.1104/pp.114.256552

Figure 5.

Figure 5.

In vitro binding assay of dsDNA and the STOP1 protein using an AlphaScreen system. A, In vitro-translated STOP1 protein labeled with the accepter beads of the AlphaScreen system was incubated with the 30-bp dsDNA. B, Relative AlphaScreen signals were calculated as the ratio of AlphaScreen signals of the reactive probe (biotin labeled) to those of the nonreactive probe (nonbiotin labeled) in the presence of the labeled STOP1 protein and streptavidin-coated donor beads. Values are the mean ± sd (n = 3). Different letters above the bars indicate a significant difference (P < 0.05, Tukey’s test). C, Competitive assays of the probe 3 region with the single nucleotide mutagenized probes. The reactive probe 3 (see B) was incubated with the labeled STOP1 protein in the presence of nonlabeled probe 3 or the probe that carried a single-nucleotide mutation. Relative values ± sd (n > 3) were calculated as the ratio of the value obtained in the absence of the competitor (AC). Asterisks indicate a significant difference larger than the relative AlphaScreen signals of nonreactive probe 3 (Student’s t test; *P < 0.05, **P < 0.01).