Six days after seeding, growth medium of human macrophages was replaced with fresh RPMI 1640 containing 2% human AB serum, penicillin (100 U/ml), streptomycin (100 mg/ml), and Lglutamine (2 mM). Cells were then incubated for one day at 37uC in a humidified, 5% CO 2 environment. Cells from donors D1, D2, and D3 were infected with either mock virus preparation or ZEBOV at a multiplicity of infection (MOI) of 10. Cells of donors D4, D5, and D6 were infected with ZEBOV at an MOI of 100, mock virus preparation, ,100 particles/cell VLP VP40-GP , ,100 particles per cell VLP VP40 , mock VLP (plasmid only), ,100 latex particles/cell, or 10 ng/ml of lipopolysaccharide (LPS). Cell supernatants were removed from the cells 1 h or 6 h post infection, and RNA from cells was purified using the RNeasy Mini Kit (QIAGEN, Valencia, CA) according to the manufacturer's instructions. The analysis of purified RNA from 10 MOI was analyzed by DNA microarrays as outlined below. The RNA obtained after infection with 100 MOI was subjected to quantitative real-time RT-PCR analysis only.