Figure 2

Ube3A^loxp+/camkIIa-cre⁺ mice have increased Ephexin5 expression in the hippocampus and Ephexin5 dependent deficits in NPP. (A) 8 week old TdTomato cre reporter-positive, camkIIa-cre⁺ tissue was stained for TdTomato (red) with Hoechst (DAPI -blue) for nuclei labeling. On the left is a 10× image of one coronal section indicating strong hippocampal and some cortical expression. On the right are images of DG and CA1, indicating nearly 100% recombination in the excitatory cell layers. Scale bar = 10 µm (B) Ephexin5 expression levels are elevated in hippocampus from ube3A^loxp+/camkIIa-cre⁺ mice. Quantification of Ephexin5 and UBE3A signal is normalized to Actin signal and compared to ube3Aloxp⁺/camkIIa-cre^- controls. Arrow indicates Ephexin5 band. Data are presented as mean ± SEM (n = 6 for ube3A^loxp+/camkIIa-cre⁺, n = 5 for ube3A^loxp+/camkIIa-cre^-). * p < 0.05 (unpaired Student’s t-test). (C) Latency for WT C57Bl/6J mice, with or without camkIIa-cre dependent ube3A removal, to fall off the rotating rod, measured in seconds. (D) Object investigation in the NPP test in 3 month-old C57Bl/6J mice, with or without camkIIa-cre dependent ube3A removal in a WT or E5^−/− background. Shown is the percent time spent investigating each object. Data are presented as mean ± SEM. *p < 0.05 (two-way ANOVA) compared to the stationary objects (S1 and S2) with post-hoc Bonferroni multiple comparisons test. See also Supplementary Fig. 2. Sample size (n) reported in Materials and Methods. Degrees of freedom and exact p values are reported in Supplementary Table 2 and Supplementary Table 3. Behavior analysis used ODlog (v 2.5 http://www.macropodsoftware.com/odlog/).