One week after the tumor injection, the tumor-bearing mice were treated intraperitoneally with 2.5 mg/kg GD5 or 3.5 mg/kg DOX. After 2 and 4 h, the spleens were collected, and the splenocytes were prepared according to the manufacturer's instructions (BioLegend). Approximately 1 × 106 cells were stained with surface antibodies and analyzed by FACSCalibur flow cytometry (BD Biosciences). The antibodies including anti-CD11c-Alexa488, -CD86-PerCP/Cy5.5, -CD80-Alexa647, -CD3-Alexa488, -CD19-Alexa647 and -CD69-PerCP/Cy5.5 were purchased from BioLegend.
Sera samples were collected at 2, 4 or 24 h from the same administrated mice and stored at -20°C. ELISA was performed using a Ready-SET-Go!® ELISA kit (eBioscience).
Sera samples were collected at 2, 4 or 24 h from the same administrated mice and stored at -20°C. ELISA was performed using a Ready-SET-Go!® ELISA kit (eBioscience).