Figure 8: Contribution of NRSF in the altered HPA axis during acute and chronic stresses. SD rats underwent traumatic stress, were subjected to predator exposure following trauma (trauma+PSS) for 14 days, and NRSF was over-expressed by adeno-virus: (A) hypothalamus was sonicated using a tissue extraction reagent supplemented with a protease inhibitor cocktail, the homogenate was centrifuged (10 min, 14,000 g, 4°C) and the supernatant collected; (B and C) cardiac blood was centrifuged (10 min, 14,000 g, 4°C), and CRH release, ACTH and CORT were measured using a competitive immunoassay. Total protein was quantified using a Bradford assay. (D) Nucleus GR expression was determined by western blot analysis. Data were calculated, each value represents the mean ± SEM of three independent experiments. *p<0.05 vs. control, #p<0.05 vs. day 1/3.