Western blot analysis

The Postsynaptic Density Protein 95 (PSD-95) antibody (95 kDa, 1:1,000, Cell Signaling, Danvers, MA) and synaptophysin (SVP) antibody (38kDa, 1:1,000, Abcam, Cambridge, MA) were used in the Western blot analysis to detect the levels of PSD-95 and SVP, respectively. The quantification of Western blot was performed as described in our previous studies [16,37]. Specifically, signal intensity was analyzed using image analysis program Quantity One (Bio-Rad, Hercules, CA). The Western blots were then quantified in two steps. First, we used β-actin levels to normalize protein levels (e.g., determining the ratio of PSD-95 to β-actin amount) and control for loading differences in the total protein amount. Second, we presented the changes in protein levels from the mice undergoing anesthesia/surgery as a percentage of those in the mice from control group. 100% changes of protein levels refer to control levels for the purpose of comparison with experimental conditions.