Tail vein injections
This animal study was conducted in accordance with the rules and regulations of the IACUC at the Department of Laboratory Animal Science, Fudan University (Shanghai, P.R. China). Lewis lung carcinoma (LLC) cells were collected and resuspended in serum-free medium. C57/BL6 mice were injected through the tail vein with 1×106cells in 0.1 mL. Fourteen days after injection of cells, the mice were subjected to PET scanning. Mice were euthanized by pentobarbital sodium, and all lungs were dissected or further imaged by fluorescence stereomicroscopy and then were fixed in Bouin’s solution, and the number of metastatic nodules on the lung surface was counted.
Subcutaneousinoculation of tumors in mice
This animal study was conducted in accordance with the rules and regulations of the IACUC at the Department of Laboratory Animal Science, Fudan University (Shanghai, P.R. China). Tumor cells at a density of 1×106in 0.1mL serum-free media were injected subcutaneously into the right flank of mice. The mice were monitored daily for the development of visible tumors. Once a tumor was clearly visible, it was calipered three times each week, and the volume was estimated using the formula V = (LW2)/2 where V is volume, L is longest diameter, and W is shortest diameter. Mice were euthanized by pentobarbital sodium, and the tumors were removed, some were fixed in 4% paraformaldehyde, and the other tumors were lysed for Western blotting.