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. 2020 Jun 9;10:9011. doi: 10.1038/s41598-020-65847-8

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Summary of animals, experiments and timeline. Whole groups (n = 10) adult mice (2 months old, Balb/c line) weighting 25-27 g on the test day was conducted behavioral test novel object, social behavioral test, Marble burying test and tail suspension test respectively. *p < 0.05, **p < 0.001, ****p < 0.0001. After behavioral test, whole groups of mice (n = 5) was sacrificed and blood, hippocampus and sperm samples were harvested. VPA (Valproic acid) Cc2d1a group G1 = Cc2d1a^+/− x Cc2d1a^+/+, G2 = Cc2d1a^+/− x Cc2d1a^+/−, control = Cc2d1a^+/+ x Cc2d1a^+/+ crosses are presented. Novel object recognition (NOR) test. Time spent with familiar versus novel objects (see Materials and Methods). Object recognition is measured as the difference in the time spent with the familiar versus the new objects. (A) Valproic acid-treated males (500 mg/kg dose). (B) Cc2d1a group, offspring from G1 = Cc2d1a^+/− x Cc2d1a^+/+, G2 = Cc2d1a^+/− x Cc2d1a^+/−, control = Cc2d1a^+/+ x Cc2d1a^+/+ crosses are presented. Controls for all groups are normal wild type Cc2d1a^+/+ Balb/c, that never been crossed with the mutants mice. Social interaction test To determine whether the interactions varied between groups of mice, the mice were placed in the center of a cage divided into three compartments, where chamber A contained another mouse, and chamber B was empty (see Materials and Methods). They were filmed for 5 minutes to calculate the time that they spend close to the empty compartment or to the compartment occupied by another mouse. (C)Valproic acid-treated males (500 mg/kg dose). (D) Cc2d1a group males, offspring from G1 = Cc2d1a^+/− x Cc2d1a^+/+, G2 = Cc2d1a^+/− x Cc2d1a^+/−, and control = Cc2d1a^+/+ x Cc2d1a^+/+ crosses are presented. Controls for all groups are normal wild type Balb/c, that never been crossed with the mutants mice. Tail suspension test The test consists of short-term suspension of mice by their tail (six minutes here). They were filmed and analyzed for immobility posture (see Materials and Methods). (E) Valproic acid-treated males (500 mg/kg dose). (F) CC2d1a group males, offspring from G1 = Cc2d1a^+/− x Cc2d1a^+/+, G2 = Cc2d1a^+/− x Cc2d1a^+/−, and control = Cc2d1a^+/+ x Cc2d1a^+/+ crosses are presented. Controls for all groups are normal wild type Cc2d1a^+/+, Balb/c, that never been crossed with the mutants mice. Glass marble burying test Repetitive action was evaluated via the marble burying test (see Materials and Methods). Twenty glass marbles were placed on the surface of clean bedding, and the number of marbles buried in 30 minutes was scored. (G) Valproic acid-treated males (500 mg/kg dose). (H) CC2d1a group males, offspring from G1 = Cc2d1a^+/− x Cc2d1a^+/+, G2 = Cc2d1a^+/− x Cc2d1a^+/−, control = Cc2d1a^+/+ x Cc2d1a^+/+ crosses are presented. Controls for all groups are normal wild type Cc2d1a^+/+, Balb/c, that never been crossed with the mutants mice.