Western blots of NT-3, BDNF, and NGF were studied according to the method described in [18] . The rat cortex was homogenated and the protein content of the supernatant was estimated with an absorbance of 560 nm by using a bicinchoninic acid protein assay reagent kit (Pierce, Rockford, IL, USA). After electrophoresis and protein transfer to a polyvinylidene fluoride membrane, the membrane was stained with primary and secondary antibodies. After these procedures were complete, the membrane was reacted in a chemiluminescence reagent (Western LightningTM, Chemiluminescence Reagent Plus, PerkinElmer Life Sciences, Inc., MA, USA). It was then exposed to a high-performance chemiluminescence film (Amersham Biosciences, England, UK) to measure the optical intensity.
A fluorescence imaging system (Vilber Lourmat, France) was used to quantify the immunoreactivity and western blots according to the method described in [19] (link). The background optical intensity of the area near the examined cortical area and the immunoreactive band was measured. The optical intensity ratio of the ischemic cortex to that of the sham operation was determined after subtracting the background intensity. The means ± SDs of these values were calculated and were statistically compared at each time point between the 2 groups. Eight rats were included at each time point for the analysis.
A fluorescence imaging system (Vilber Lourmat, France) was used to quantify the immunoreactivity and western blots according to the method described in [19] (link). The background optical intensity of the area near the examined cortical area and the immunoreactive band was measured. The optical intensity ratio of the ischemic cortex to that of the sham operation was determined after subtracting the background intensity. The means ± SDs of these values were calculated and were statistically compared at each time point between the 2 groups. Eight rats were included at each time point for the analysis.