Figure 4.

GSK3β interacts and regulates FoxO3a in mouse cerebral cortex. (A) The cerebral cortex from control (C) and IES (S)-treated mice were dissected on post-IES day-3 and day-8. Protein homogenates were immunoprecipitated with anti-FoxO3a or anti-GSK3β, and immunocomplex and homogenates (input) were immunobloted for FoxO3a and GSK3β. Data is expressed as the ratio of the protein in the immunocomplex (Co-IP) to total lysate. Mean ± SEM, n=4-6/group, *p<0.05 in Student’s t-test when IES-treated mice were compared to control mice. (B-D) Control and IES-treated mice received daily i.c.v. infusion of saline or the GSK3 inhibitor BIP-135 (0.08 nmol). (B) LH behavior and social interaction were tested on post-IES day-1, 2 hr after GSK3 inhibitor treatment. Mean ± SEM, n=6/group, *p<0.05 in Student’s t-test. (C) After 3 doses of BIP-135 on post-IES day-1, 2, and 3, protein homogenates from the cerebral cortex were immunprecipitated for FoxO3a and the immunocomplexes were immunobloted for FoxO3a and GSK3β. Data is expressed as the ratio of the protein in the immunocomplex (Co-IP) to total lysate. Mean ± SEM, n=6/group, *p<0.05 in two-way ANOVA with post-hoc analysis. (D) After BIP-135 on post-IES day-1, 2, and 3, nuclear proteins from the cerebral cortex were immunobloted for FoxO3a. Data is expressed as % control (no IES). Mean ± SEM, n=6/group, *p<0.05 in Student’s t-test.