Fig. 3

Evaluation of cellular uptake, antitumor activity and mechanism of ACaT in vitro. A CLSM images of SKOV3 cells after treatment with ACaT/6-FAM for 0, 1, 2, 4 and 24 h. Scale bar, 30 μm for the three columns on the left and 10 μm for the enlarge column. B Quantitative cell cycle analysis of different phases in different treatment groups. C Top 10 KEGG pathways enrichment analysis bubble-plot. Cells were treated with 100 mg/L of ACaT for 24 h before mRNA sequencing. Black arrow indicated cell apoptosis and cell cycle pathway. D Dual AO/EB fluorescent staining of SKOV3 cells after treatment with different concentrations of ACaT for 48 h (green for living cells and red for dead cells). Scale bar, 150 μm. E Cell viability of SKOV3 treated with ACaT at different concentrations for 24, 48 and 72 h. F Western blots for Bax, Bcl-2, and cleaved caspase-3 after NaALN (850 μM), THZ1 (0.5 μM), ACaT (100 mg/L) treatment for 24 h in SKOV3 cells. G Scratch assay for cell migration. Cell migration was recorded at 0 and 24 h after scratching. Cells were treated with NaALN (200 μM), THZ1 (0.01 μM), ACaT (6.25 mg/L) at the beginning of the experiment (yellow dotted boxes marked the scratch edges)