(a) Sequences and modifications of NBOP-17Dz-DQ, DEACM-17Dz-mod-DQ,
17S-FQ, and 17S-mod-CQ used in the cellular study. C is the Cy5 dye.
Other labels are the same as those used in Figure 4. The binding arms of DEACM-17Dz-mod-DQ were
modified to recognize the substrate 17S-mod-CQ and to eliminate cross
talk with NBOP-17Dz-DQ + 17S-FQ. (b) Confocal images of HeLa cells
transfected with PS-17Dz-DQ, PS-17Dz-mod-DQ, 17S-FQ, and 17S-mod-CQ
under different assay conditions. Both PS-17Dz-DQ and PS-17Dz-mod-DQ
were active in cleaving their cognate substrates to release fluorescent
signals. (c) Sequential activation of NBOP-17Dz-DQ and DEACM-17Dz-mod-DQ
by visible and UV light and global decaging of both DNAzyme sensors
by UV light. Green and red fluorescence arose from the cleavage of
17S-FQ and 17S-mod-CQ, respectively, in the presence of active cognate
DNAzyme sensors. Scale bars = 100 μm. For error bars, n = 3. Conditions for light irradiation: 365 nm at 56 mW/cm2 for 5 min and 470 nm at 20 mW/cm2 for 10 min.
(d) Flow cytometer analyses of samples shown in (b) and (c).