In the physiological environment: The working solution for each DNA nanostructure was adjusted to 20 nM and then mixed with DMEM (supplemented with 20% FBS) at a 1:1 volume ratio. All samples were immediately incubated at 37°C for 2, 4, and 6 h.
In the low-pH environment: To mimic the cleavage situation of DNA nanostructures in cellular lysosomes, DOS, DTU, and DTE were treated in the low-pH environment (pH = 5) for different periods at 37°C.
Right after incubation, different DNA samples were subjected to 10% native polyacrylamide gel electrophoresis (PAGE) at 100 V for 1 h (gel prepared in 1 × TBE buffer supplemented with 10 mM MgCl2). After the electrophoresis, the gels were carefully recovered and incubated with 1 h in the staining solution (3 × SYBR Safe was dissolved in 1 × TBE buffer). Finally, the stained gels were exposed using the Tanon 4600SF multifunctional imaging system (Tanon, China).
In the low-pH environment: To mimic the cleavage situation of DNA nanostructures in cellular lysosomes, DOS, DTU, and DTE were treated in the low-pH environment (pH = 5) for different periods at 37°C.
Right after incubation, different DNA samples were subjected to 10% native polyacrylamide gel electrophoresis (PAGE) at 100 V for 1 h (gel prepared in 1 × TBE buffer supplemented with 10 mM MgCl2). After the electrophoresis, the gels were carefully recovered and incubated with 1 h in the staining solution (3 × SYBR Safe was dissolved in 1 × TBE buffer). Finally, the stained gels were exposed using the Tanon 4600SF multifunctional imaging system (Tanon, China).
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