Fig. 1

Standardization of the tumor model and its tentative mode of action The animals were vaccinated for two times before injected tumor cells during pre-experimental stage. Panel (a), shows the tumor growth after day 0, 10 and 30, as a result of subcutaneous injection of CT26.WT tumor cells into the neck of BALB/c mice. Panel (b), depicts the efficacy of cytokine adjuvant vaccine, where each group with 5 mice was vaccinated twice at interval of 7 and 3 days, before subcutaneous injection of CT26.WT cells. Panel (c), shows the comparison between male and female mice in terms of tumor uniformity. Panel (d), represents the standardization of inactivated antigen for effective anti-tumor response. M stand for million, * stand for Optimization. Panel (e), represents the effect of combination application compared to single group by observing survival period, which each group with 15 mice was vaccinated twice before subcutaneous injection of CT26.WT cells. *denotes p < 0.05, ***denotes p < 0.001. Panel (f), The experiment timeline showing the times of vaccinated. In order to establishing the tumor recurrence model stimulating the clinical status, the experiment was divided into prevention stage and therapeutic stage. The Prevention group was vaccinated vaccine for two times in prevention stage, the Treatment group was vaccinated vaccine for four times include prevention stage and therapeutic stage, and the Tumor group vaccinated inactivated antigen for control. All groups were inoculated tumor cells at the same time. Tumor inoculation was the initial point (day 0)