Figure 2

Two-dimensional gel electrophoresis maps of protein extracts prepared from PM of control (A) and morphine-treated (B) rats. Protein samples (400 μg for Silver staining; 2 mg for CBB staining) were separated in the first dimension on pH 3–11 IPG strips. For resolution in the second dimension, SDS-PAGE was performed in 10% w/v acrylamide/0.26% w/v bis-acrylamide gel (silver staining) or in 12.5% w/v acrylamide/0.0625% w/v bis-acrylamide gel (CBB staining). The stained 2D gels were scanned in an imaging densitometer and quantified by PDQuest software. The process of quantification of the difference between morphine-treated (+M10) and control (−M10) samples included spot detection, gel matching and spot quantification. Master gel was constructed for each group (+M10) or (−M10) as a synthetic image that contains the spot data from all the gels in the MatchSet. At least four replicates were performed for each group/sample. All matched and unmatched spots were then checked in a manual manner. Protein levels altered at least two-fold were taken into consideration. About 500 protein spots totally were recognized by CBB staining by PDQuest analysis. Proteins 1–18 with an altered mobility in (+M10) versus (−M10) samples were excised from in CBB-stained gels and identified by LC-MS/MS as described in methods. Left panels, Silver staining. Right panels, Coomassie staining.