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. 2018 Dec 12;47(3):1097–1109. doi: 10.1093/nar/gky1245

Figure 5.

Figure 5.

(A) Illustration of feedback circuit. The fluorophore FAM and the quencher BHQ are functionalized at either end of substrate strand R2. (B) Native PAGE analysis of feedback-circuit products. Lane 1, DNA complex Z3/T5/R2; lane 2, Unit1 complex Z3/T5/R2/P1; lane 3, products of Unit1 triggered by input I6; lane 4, Unit2 complex Z4/T1/R4; lane 5, products of Unit2 in presence of input I6; lane 6, products of Unit2 triggered by input I1; lane 7, mixture of Unit1 complex and Unit2 complex; lane 8, products of feedback circuit consisting of Unit1 and Unit2 triggered by input I6. (C) Time-dependent normalized fluorescence changes (ΔF/MaxΔF) during 10-h reaction process. The sample interval was 6 min. As baselines, curve (1) demonstrates the leakage level of Unit1 and curve (2) demonstrates the leakage level of feedback circuit in absence of trigger I6. Curve (1′)-(3′) demonstrate the Unit1 responses at different concentrations of I6 as 0.05, 0.1 and 0.15 μM, respectively. All data represent the average of three replicates. Error bars represent one standard deviation from triplicate analyses. (D) Time-dependent normalized fluorescence changes (ΔF/MaxΔF) during 10-h reaction process. The sample interval was 6 min. Curves (1)–(3) demonstrate the feedback-circuit responses at different concentrations of I6 as 0.05, 0.01, 0.15 μM, respectively. Curves (1′)–(3) demonstrate the Unit1 responses at different concentrations of I6 as 0.05, 0.1 and 0.15 μM, respectively. All data represent the average of three replicates. Error bars were not plotted to avoid curves overwriting each other. (E) Fluorescence signal analysis in form of bars. The columns 1–3 correspond to the normalized fluorescence changes (ΔF/MaxΔF) of the Unit1 triggered by input I6 at 0.05, 0.1, 0.15 μM, respectively, after 10 h. The columns 4–6 correspond to the normalized fluorescence changes (ΔF/MaxΔF) of feedback circuit triggered by I6 at 0.05, 0.1, 0.15 μM, respectively, after 10 h. And the relative fluorescence increase percentage (F2/F1%) was labeled at the top of bars in columns 4–6.