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. 2013 Sep 3;4:2393. doi: 10.1038/ncomms3393

Figure 6. Simultaneous and symmetrical control of two outputs in a cell by using ON and OFF switches.

Figure 6

(a) The behaviour of ON and OFF switches responsive to a single input (L7Ae). Two reporter proteins, EGFP and enhanced cyan fluorescent protein (ECFP), were produced from ON and OFF switches, respectively. Two OUTPUT plasmids expressing either an ON or an OFF switch (100 ng each) were transfected into HeLa cells together with 20 ng of cognate and 480 ng of noncognate INPUT plasmids. The fluorescent signals were normalized by those of the switches containing Kt in the absence of L7Ae. (b) Control of two switches under two input proteins. Kt or dKt in ON switches were replaced with Fr15 or dFr15 (defective Fr15). Two OUTPUT plasmids were transfected together with L7Ae- (20 ng) and/or S15- (480 ng) expressing plasmids. In this experiment, a viral nucleocapside protein was used as a noncognate RNA-binding protein. The fluorescent signals were normalized by those detected in the absence of both L7Ae and S15. Transfected cells were analysed using FACSAria. Bars and error bars represent the mean and s.d., respectively, of three replicates.