4.3.3 EdU incorporation assay
EdU incorporation assay were performed using BeyoClick™ EdU Cell
Proliferation Kit with Alexa Fluor 488 (Beyotime). Cells
(4×104 cells/mL) in 24-well were treated with ANVs or
AIE@ANVs for 24 hours, then incubated with EdU (20 mmol/L) for 2 hours.
The cells were fixed with 4% paraformaldehyde for 20 minutes at room
temperature, and washed with PBS (containing 3% BSA). Then, the cells
were treated with Triton X-100 (0.3% in PBS) for 10 min. After washing
again with PBS, the cells were incubated with 100μl click additive
solution (containing CuSO4 and Azide 488) at room
temperature for 30 min. Nucleus was stained by 1× Hochest 33342 at room
temperature for 10 min. After washing with PBS, the cells were observed
by fluorescence microscopy.