SDS-PAGE was performed as described in our previous work (9). A 4 to 20% gradient gel (Bio-Rad) was used, and 0.5 μg of each Dox conjugate was loaded with or without reduction with 10 mM dithiothreitol. After electrophoresis, SimplyBlue SafeStain (Thermo Fisher Scientific) was used for gel staining according to the manufacturer’s instruction. ChemiDoc XRS+ system (Bio-Rad) was used for image acquisition.

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