Binding assay of [³H]15d-PGJ₂ to subcellular fractions.

<p>(A) Proteins fractionated to plasma membranes, nuclear, cytosol or microsome from cortical neurons were incubated with 10 nM [³H]15d-PGJ₂ at 25°C for 1 h in the absence or presence of 10 µM unlabeled 15d-PGJ₂. Total binding, nonspecific binding and specific binding were hatched columns, open columns and closed columns, respectively. Data are expressed as means (n = 2). (B) Effects of 15d-PGJ₂ and its precursors on the binding of 10 nM [³H]15d-PG J₂ to cortical neurons (open columns) and BSMC (closed columns). Plasma membranes (10 µg/protein) were incubated with [³H]15d-PGJ₂ at 4°C for 24 h in the presence of unlabeled PGD₂, PGJ₂, Δ¹²-PGJ₂ or 15d-PGJ₂ at the indicated concentrations. The control value of [³H]15d-PGJ₂ binding in cortical neurons and BSMC were 2523 cpm and 1309 cpm, respectively. Data are expressed as means ± S.E.M. (n = 4). *P<0.05, **P<0.01, compared with control by ANOVA followed by Dunnett's test.