Distribution of the myc-tagged 14-3-3ζ transgene in mouse brain.

<p>(A) Western blot analysis (<i>n</i> = 1 per lane) of microdissected brain regions from wild-type (wt) and 14-3-3ζtg mice immunoblotted with antibodies against the myc tag confirm expression of the transgene in CA subfields, dentate gyrus (DG), cortex (Cx), cerebellum (Cb), striatum (Str) and brain stem (BS). (B) 14-3-3ζ immunostaining in hippocampal tissue sections showing higher immunoreactivity (IR) in 14-3-3ζtg compared to wt mice. Panels to the far right show sections stained for 14-3-3γ revealing normal neuronal distribution and level between wt and 14-3-3ζtg mice. (C) Representative fluorescence immunostaining of the CA1 subfield showing (top) myc (red) with NeuN (green) co-localization and (bottom) myc (green) with GFAP (red) confirming mainly neuronal expression of the transgene. (D) Immunoblots of SJL mouse hippocampal fractions (<i>n</i> = 1 per lane) for the select presence of markers of the nucleus (Nuc), mitochondria (Mito), cytoplasm (Cyto) and microsome-containing ER fraction (Micro). (E) Western blots from pools of 14-3-3ζtg mouse hippocampi (<i>n</i> = 1 per lane) show the presence of various isoforms in different compartments. Note, endogenous 14-3-3ζ and myc-14-3-3ζ are similarly distributed in each fraction. (F) Protein levels of various 14-3-3 isoforms in wt and 14-3-3ζtg mice hippocampal subfields (<i>n</i> = 1 per lane). (G) Real-time PCR measurement of ζ, γ and ε 14-3-3 isoform levels in wt and 14-3-3ζtg mice (<i>n</i> = 3 per group). *<i>p</i><0.05 compared to wt. Scale bars in B, C, 160 µm.