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. 2018 Oct 9;12:341. doi: 10.3389/fncel.2018.00341

Figure 2.

Figure 2

Generation and characterization of global Shank3Δ14–16 and Viaat-Cre;Shank3Δ14–16 mice. (A) conditional knockout (cKO) strategy for exons 14–16 of the Shank3 gene in mice. Ankyrin, ankyrin repeats; SH3, src homology 3 domain; PDZ, PSD-95, Dlg, ZO-1 domain; Pro-rich, proline-rich region; SAM, sterile alpha motif; Ex, exon; Frt, flippase recombinase target; loxP, locus of X-over P1. (B) Polymerase chain reaction (PCR) genotyping of global Shank3Δ14–16 and Viaat-Cre;Shank3Δ14–16 mice. Note that the primer set targeting exons 13 and 14 generates a PCR band (276 bp) in the wild-type (WT) allele, but not in the Shank3Δ14–16 (del) allele, whereas the primer set targeting exons 13 and 17 generates a PCR band (1159 bp) in the Shank3Δ14–16 (del) allele, but not in the WT allele. (C) Immunoblot analyses of Shank3 protein splice variants in WT and Shank3Δ14–16 mice (13 weeks; male) using a Shank3-specific antibody (#2036); the target region is indicated in panel A as a red bar. Th, thalamus; Str, striatum; Hpc, hippocampus; Ctx, cortex. (D) Reduced levels of Shank3 protein variants in different brain regions of Viaat-Cre;Shank3Δ14–16 mice (12 weeks; female). Total brain lysates were analyzed by immunoblotting using a Shank3-specific antibody (#2036). cKO band signals normalized to α-tubulin were normalized to those from WT mice. Data are shown as mean ± SEM. n = 3 pairs (WT, cKO), **P < 0.01, nd, not detectable, ns, not significant and one sample t-test.