Figure 4.

Cell cytotoxicity assay of drug-loaded micelles on dog bladder cancer. K9TCC-Pu-In cells were seeded and cultured overnight before being treated with various concentrations of nontargeting or targeting micelles loaded with daunorubicin (DNR) for two hours at 37°C. Cells treated with empty micelles and free DNR under the same conditions served as controls. Cells were then washed and cultured with complete culture medium for 72 hours and the cell viability was evaluated using the WST-8 assay according to the manufacturer’s protocol. The absorbance was detected using an enzyme-linked immunosorbent assay reader. This experiment was performed in triplicate and repeated three times.

Notes: *P = 0.02 for free DNR versus PLZ4 micelles and 0.04 for free DNR versus nontargeting micelles. **P = 0.001 for free DNR versus PLZ4 micelles; 0.01 for free DNR versus nontargeting micelles; and 0.04 for nontargeting versus targeting micelles.