Transwell migration was monitored using 96-well BD Fluoroblock plates with 8 µm filters (Corning, Inc., Corning, NY). Cells were harvested and suspended at density of 2 × 106 cells/ml in migration media (L-15 + 1% FBS), and a volume of 75 μl cell suspension was placed on top of the filter inserts. Where indicated, IL-10R neutralizing antibody was added at a concentration of 30 μg/ml. The bottom wells were loaded with the indicated concentrations of EGF in the presence of conditioned medium from mock or HCMV-infected fibroblasts (96 h post infection) in a total volume of 235 μl. After 5 h at 37 °C, cells that traversed the filter and entered the lower chamber were quantified by the addition of Cell Titer Glo (Promega, Madison, WI) using a Turner Veritas luminometer. For invasion, 96-well matrigel-coated BD Fluoroblock transwell invasion plates (Corning) were used. Invasion plates were re-hydrated with warm media at 37 °C for 3 h and then 75 μl cell suspension loaded onto the hydrated filters as described above. Where indicated, 10 μM Stat3 inhibitor was included with cells in the top chamber; cmvIL-10, hIL-10 or conditioned medium was present in both chambers. The bottom plates received the indicated EGF concentrations, and then transwell system was incubated for 22 h at 37 °C with atmospheric CO2. At harvest, cells that had degraded the matrigel and entered the lower chamber were quantified by the addition of Cell Titer Glo as above.