LC-MSMS of Brucella infected RAW264.7 cells

The RAW264.7 cells (2x105 cells/ml) were seeded in 100 mm cell culture dishes in DMEM containing 10% FBS. When the cell density reached 70%, cells were infected with B. abortus A19 (MOI=200) for 4 h, and the cell medium then was changed to DMEM containing 10% FBS with gentamicin (50 ng/μl) for 1 h. The medium subsequently was changed to DMEM containing 10% FBS with gentamicin (25 ng/μl) for 24 h at which time cells were harvested and placed at -80°C. Subsequently, the cells lysates were analyzed by protein liquid chromatography-mass spectrometry (LC-MS/MS)