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Prostaglandin 2 (PGE2) level assay
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The levels of secreted PGE2 in the cell supernatants were quantified using an prostaglandin E2 enzyme-linked immunosorbent assay (ELISA) kit (Cayman Chemical Company, Ann Arbor, MI, USA) as described in the manufacturer’s instructions and presented as pg/mL. Breafly, neurospheres were treated in presence or absence of NS398 (10 µM, 100 µM, and 200 µM) for 48 h; cell culture supernatants were assayed at a final dilution of 1:10 for U87MG and not diluited for T98G cultures. The least detectable PGE2 concentration was 7.8 pg/mL.
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