Figure 5. Altered expression of Ube3a and Shank3 in the amygdala ofUba6^NKOmice.

(A) Immunoblotting of amygdala extracts from Uba6^NKO mice at 3 months of age with the indicated antibodies. Tubulin was used as controls. Extracts from two mice of each genotype were employed.

(B,C) qPCR analysis of Ube3a and Shank3 mRNA from the amygdala of two control and Uba6^NKO mice. Data are mean ± SEM, with n=3 independent experiments. (D,E) Increased abundance of Ube3a in Uba6^NKO mice correlates with loss of Arc expression in the amygdala in two animals (D), but without alterations in Ubr2 or Mdm2 (E). (F-H) Uba6 is required for hippocampal neuron viability in dissociated cultures in vitro. Hippocampal cells from E16.5 embryos were depleted with shRNAs targeting Uba6 for 5 or 12 days and the presence of Uba6 at day 5 (F) or the presence of neurons at day 5 (G) or 12 (G,H) determined using anti-MAP2 antibodies. For quantification, the number of neurons in 12 random 360,000 μm² fields were determined from duplicate (5 h) or triplicate (12 h) experiments. Error bars in panel G are means of ± SEMs (n=3 independent experiments).