Figure 2. Characterization of the GFP-dependent transcription system.
(A) Schematic of Gal4-based T-DDOGs. (B) GFP-dependent activation of UAS-luc2 by Gal4-GBP6VP16-GBP1 and Gal4-GBP2VP16-GBP7. n=9. (C) Gal4-GBP6VP16-GBP1 strongly activated UAS-tdT in the presence of GFP. Mutation of GBP1-binding residues in GFP (GFPmG1) abolished tdT activity. Scale bar, 10μm. (D) Specificity of T-DDOGs for different fluorescent proteins. n=9. *, p<0.001. (E) Activity of Gal4-GBP6VP16-GBP1 in response to varying amount of transfected GFP plasmids. The transfected DNA amount was kept constant among conditions, with CAG-mCherry (bottom panel) acting as a filler plasmid to compensate for reduction in GFP (top panel) plasmids. Panels show representative GFP and mCherry fluorescence in single cells for each corresponding data point below. n=6. Plots are mean +/− SD. See also Figure S1, S2 and Table S2.