Determination of μ-OR in PNS prepared from FBC of experimental groups (+M10) and (─M10).

<p>PNS fractions (20 μg protein per lane) prepared from morphine-treated (+M10) and control (— M10) rats were resolved under non-dissociated (— DTT) or dissociated (+DTT) conditions by 1D-SDS-PAGE in 10% w/v acrylamide/0.26% w/v bis-acrylamide (<b>A, C)</b> or in 4–12% NuPAGE gels (<b>B, D</b>); μ-OR was recognized by C-terminus-oriented Ab C-20 (sc-7488-R). When resolved under non-dissociated conditions (<b>A</b> and <b>B</b>), Ab C-20 distinguished the single protein with M_w ≈ 60 kDa. The average intensity of this immunoblot signal was not significantly different when compared in control (— M10) and morphine-treated (+M10) PNS samples (NS, p>0.05). Resolution under dissociated conditions (<b>C</b> and <b>D</b>) revealed the presence of multiple protein bands exhibiting a wide range of M_w ≈ 20–90 kDa. These immunoblot signals were also unchanged by morphine (NS, p>0.05). The results (upper columns) represent the average signal of five immunoblots, each performed with four control + four morphine-treated samples of PNS ± SEM. 100% on y-axis represents the average intensity of a given immunoblot signal determined in PNS prepared from control, (─M10) rats. The significance of difference between the control and morphine-treated PNS samples was analyzed by Student´s <i>t</i>-test using GraphPad<i>Prizm4</i>. In the lower panels, typical immunoblots are presented.