The cells and brain tissues were harvested at the end of the experiment and were subjected to Western blot analysis, as described by Xie et al.25 Antibodies A8717 (1:2000; Sigma, St Louis, Missouri), C66 (1:1000; generous gift of Dora Kovacs, PhD, at Massachusetts General Hospital and Harvard Medical School), and anti–β-actin (1:5000; Sigma) were used to visualize FL-APP (110 kDa), APP-C83 (12 kDa), APP-C99 (10 kDa), and β-actin (42 kDa), respectively. A caspase-3 antibody (1:1000; Cell Signaling Technology Inc, Beverly, Massachusetts) was used to recognize the caspase-3 fragment (17-20 kDa) resulting from cleavage at asparate position 175 and caspase-3 FL (35-40 kDa). Rabbit polyclonal anti–BACE1 antibody ab2077 (1:1000; Abcam, Cambridge, Massachusetts) was used to detect the protein levels of BACE (65 kDa). The quantification of Western blots was performed in 2 steps, as described by Xie et al.25 Briefly, the intensity of signals was analyzed by using an image program from the National Institutes of Health (NIH ImageJ; Bethesda, Maryland). First, we used levels of β-actin to normalize (eg, determining ratio of FL-APP amount to β-actin amount) the levels of FL-APP, APP-C83, APP-C99, FL-caspase-3, caspase-3 fragment, BACE, and Aβ to control for the loading differences in total protein amounts. Second, we presented the changes in the levels of FL-APP, APP-C83, APP-C99, FL-caspase-3, caspase-3 fragment, Aβ, and BACE in the cells or animals treated with sevoflurane, Z-VAD, Aβ, and L-685,458 as the percentage of those in the cells or animals treated with controls. We refer to 100% caspase-3 activation, FL-APP, APP-C83, APP-C99, Aβ, and BACE in this article as control levels for the purpose of comparison with experimental conditions.