TLR agonists synergistically enhance rlipo-E7m-induced dendritic cell activation and anti-tumor activity. Cultured plasmacytoid dendritic cells (pDCs) were incubated with medium, CpG ODN (100 nM), imiquimod (10 μg/ml) or rlipo-E7m (100 nM) ± CpG ODN or imiquimod. The supernatants were collected for the detection of cytokines 24 hours after stimulation. (a) Cytokines, including IL-12p70, TNF-α and IL-10, were analyzed by ELISA to assess pDC activation. The data are presented as the means + SD of duplicate DC cultures from three independent experiments. **P < 0.01, ***P < 0.001. (b) To evaluate the anti-tumor effect of the combined formulation, tumor-bearing mice were administered a single dose of PBS, rE7m (1 μg/mouse), rE7m (1 μg/mouse) and CpG (10 μg/mouse), rlipo-E7m (1 μg/mouse) or rlipo-E7m (1 μg/mouse) and CpG ODN (10 μg/mouse) via subcutaneous (s.c.) injection 7 days after tumor cell implantation. The tumor volume was calculated using the formula length × width × width/2(mm3).