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PBS was collected, clarified by centrifugation, and subjected to PGE2 EIA kit (Cayman, Ann Arbor, MI).
Supernatants were collected, clarified by centrifugation and used to quantify cytokines as described above.
As for HDV, the tissue culture fluids were collected, clarified and then concentrated by PEG precipitation.
At 5 days post transfection, the culture supernatants were collected, clarified and then inoculated into VeroE6 cells.
The supernatants were collected, clarified by centrifugation 5 min at 600 g and 80 µl were submitted to LDH-based cytotoxicity kit (Sigma).
Two days after transfection, the medium containing HCVpp was collected, clarified, filtered through a 0.45-µm-pore-size membrane, and used for infection of Huh-7 cells.
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In contrast, in centralized hierarchies collecting, clarifying, and disseminating information about public good preference is costly, and along the information, channel toward the central government potential information leakage will be counterproductive to governmental decision-making progress, causing loss of control (Williamson 1967).
After 3 days, the cell culture supernatant was collected and clarified at 2,000 × g for 10 min at 4°C.
On day 5 post-infection, the cell supernatant was collected and clarified by centrifugation.
Briefly, Vero cells (CCL-81, ATCC, Manassas, VA) were infected with low-passage rHMPV, rHRSV, rHPIV3 or IAV at an input MOI of approximately 0.1 PFU/cell, and after 6 to 8 days, the supernatants were collected and clarified by centrifugation, and virus was collected by centrifugation on discontinuous sucrose gradients [30].
A 3 day food record will be collected and clarified.
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