Fig. 4.

Biochemical analysis of NR2D transgenic mice.A, Northern blot analysis with the RNA load controlled by reprobing the membrane with a probe for cyclophilin RNA. NR1 and NR2A levels were unaffected, but all three lines showed reduced amounts of NR2B. B, In situ hybridization of brain with a probe for NR2B. NR2B message is reduced in the transgenic mice forebrain. Scale bar, 2.5 mm. C, Immunoblot analysis of extracts from control (C) or transgenic (T) hippocampus at either 3 weeks or 2 months postnatal using antibodies againstNR1, NR2A, and NR2B.D, Immunoblot analysis of the amount of Ca²⁺-independent CaMKII activity using an antibody specific for CaMKIIα that is phosphorylated at Thr-286.Phospho-CaMKII, Thr-286 phosphorylated CaMKIIα.E, Quantitation of the relative amount of NR2B and autophosphorylated CaMKIIα in 3-week-old (3wk) or 2-month-old (2mo) transgenic hippocampi. The data were expressed as percent, with the amount in wild-type hippocampi as 100%. Values are mean ± SEM; n = 4, juvenile brain;n = 8, adult brain.