PMC:3537549 / 16923-17835 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/3537549","sourcedb":"PMC","sourceid":"3537549","source_url":"https://www.ncbi.nlm.nih.gov/pmc/3537549","text":"Production of supernatants\nFor each cat, PBMCs were resuspended in supplemented RPMI 1640 with Glutamax I (Gibco®, Invitrogen) at a concentration of 106 cells/mL in 6-well plates and stimulated immediately after isolation with 4 μg/mL ODN 2216, 4 μg/mL ODN 2243 or an equivalent volume of endotoxin-free PBS. After 24 h incubation, supernatants were harvested by centrifugation of the cultures twice at 2000 × g for 10 min, aliquoted and stored at −20°C. Large supernatant quantities were produced with PBMCs purified after one blood collection, enabling the utilization of the same supernatants for virus inhibition experiments concerning VSV, FCV, FCoV, FHV and FPV. New supernatant batches were produced for use in FeLV inhibition assays. Supernatants derived from PBMCs stimulated with ODN 2216, ODN 2243 and endotoxin-free PBS are referred to as Sup2216, Sup 2243 and Sup Neg in the text and in the figures.","divisions":[{"label":"Title","span":{"begin":0,"end":26}}],"tracks":[]}