Timing Is Critical for an Effective Anti-Metastatic Immunotherapy: The Decisive Role of IFNγ/STAT1-Mediated Activation of Autophagy
Figure 6
Therapeutic application of TLR4/TLR9 agonist complex and AG490 act synergistically to attenuate metastasis.
C57BL/6 mice were injected with B16-F10 melanoma cells (5×105) and were humanely sacrificed 14 days after tumor cell inoculation. The mice were intraperitoneally injected with the TLR4/TLR9 agonist complex (dosage and frequency stated in the legend of Fig. 1) with or without AG490 (30 mg/kg, once a day) after tumor cell inoculation. (A) Metastatic nodules were counted and summarized, and the data are the mean ± S.E. (n = 10). (B) The expression of STAT1/3 singling and autophagy-related molecules in the lung tissue. The lungs were excised and the cytoplasmic and nuclear fractions were extracted as described in the Methods. The expression of p-STAT1, STAT1, p-STAT3, STAT3, and histone H3 in nucleic extracts and IRGM1, LC3B, cleaved caspase-3, P62, PCNA, and β-actin in the cytoplasm were detected with Western blotting. Left panel is representative western blots and right panels are summary results. Data are presented as the mean ± S.E. of 5 mice per group.