Fig. 2.

Effects of anesthesia on the levels and phosphorylation/activation of various protein kinases. (A) Brain homogenates of mice anesthetized by ether inhalation for 30 sec or 5 min after intraperitoneal injection of pentobarbital were analyzed by Western blots developed with antibodies against several protein kinases and their phosphorylated/activated counterparts. Mice sacrificed without anesthesia (by cervical dislocation) were included as controls. Anti-GSK-3β (pY216) also immunoreacted with phosphorylated GSK-3α (upper band). The anti-JNK is known to detect both 46-kDa JNK1 and 54-kDa JNK2/3. Actin blot was included as a loading control. (B) Densitometric quantitation of blots as shown in (A). Actin blot was used for normalization for quantitation of the kinase levels. Each kinase blot was used for normalization of quantitation of the phosphorylated/activated form of the corresponding kinase. ^*p < 0.05 vs. controls.