Cell Apoptosis and Cell Cycle Arrest Analysis

The cell apoptosis and cell cycle arrest were carried out to investigate the mechanism of anti-proliferative effect induced by MDPAS micelles. First, HepG2 cells were seeded into 6-well plates at a density of 2.5×10⁵ cells/well, and further cultured at 37 °C overnight. Different micelles containing 1.0 μM DOX were then added into each well, and the cells were harvested and rinsed with PBS three times after the treatment for 48 h. The Annexin V-FITC/PI staining procedure was conducted in the dark for 30 min based on the apoptosis detection kit’s instructions, whereas the cells were treated with PI staining solution according to the cell cycle detection kit’s protocol. Finally, the apoptosis and cell cycle arrest induced by MDPAS micelles were analyzed through 30,000 gated cells by FACS.