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. 2018 May 28;11:177. doi: 10.3389/fnmol.2018.00177

FIGURE 1.

FIGURE 1

EEN1 KO mice are impaired in long-term spatial and contextual fear memory. (A) Domain structure of EEN1. (B) Schematic representation of the EEN1 gene locus, the KO-first, floxed and mutant alleles after homologous recombination. zptF/R and loxPF/R: primer pairs used for genotyping. neo, the neomycin resistance cassette. (C) Immunoblots of tissue lysates from mouse littermates, probed with antibodies to EEN1 and EEN2. β-Actin serves as loading control. 1, hippocampus; 2, cortex; 3, cerebellum; 4, liver. (D) No differences in the body weight of EEN1+/+, EEN1+/-, and EEN1-/- mice were detected during development (9 EEN1+/+, 11 EEN1+/-, and 14 EEN1-/-). (E–G) No effects of EEN1 KO on the performance in assays of rotarod (E and F) and Y maze (G). Data represent mean ± SEM for each group (18 EEN1+/+, 26 EEN1+/-, and 22 EEN1-/-). (H–K) No effects of EEN1 KO on the social affiliation and sociability (H and I) or social memory and novelty (J and K). Data represent mean ± SEM (9 EEN1+/+, 10 EEN1+/-, and 8 EEN1-/-). (L–S) The Morris water maze test. Shown are escape latency or traveled distance before escaping to the platform among groups in the visible-platform training (L and M), escape latency, and traveled distance before escaping to the platform in the invisible-platform training (N and O), number of crossing with the 1.5× platform area over 35 days after training and the swim trace 7 days after training in the probe test (P and Q), the swim trace and recall ability following training once again on day 35 (R and S). Red circle indicates position of the platform. Data represent mean ± SEM (9 EEN1+/+, 10 EEN1+/-, and 8 EEN1-/-), ∗p < 0.05, ∗∗p < 0.01. (T and U) Contextual fear conditioning. Shown are levels of freezing behavior after 24, 48, and 72 h from contextual fear training, and levels of freezing when animals were exposed to a novel context. Data represent mean ± SEM (9 EEN1+/+, 11 EEN1+/-, and 10 EEN1-/-), ∗∗p < 0.01