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. 2014 May 5;9(5):e95151. doi: 10.1371/journal.pone.0095151

Figure 3. Surrounding nucleotides outside of the ½ Site are crucial for binding.

Figure 3

A: EMSA was performed on mutated sequences that were generated in the second ½ site to test if variation at position, P8 (A→T), P11 (G→C) and P13 (G→T) affects binding. When P8 was changed, the faster migrating binding conformation was lost. When P11 was changed most of the slower migrating binding conformation was lost. Almost all binding was completely lost when P13 was changed. B: Luciferase reporter assays were performed to test their effect on transcription. Constructs harboring 6 copies of the mutated binding site (mutated at either P8, 11 or 13, respectively) were co-transfected with the full length Tbx1 gene. Mutation of these nucleotides affected luciferase activation. Data are presented as means ± SD; n≥3. Student's t-test, *p<0.02.