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. 2021 Jan 7;11:24. doi: 10.1038/s41398-020-01112-w

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© The Author(s) 2021

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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Fig. 4 — a Escape latency of WT, CPE-KO, and CPE-E342Q mice in Morris water maze test. WT and CPE-E342Q mice exhibited a normal learning acquisition curve during the 5-day test, and spatial memory; whereas CPE-KO mice demonstrated a deficit in spatial learning and memory. Two-way repeated ANOVA analysis followed by Tukey’s post hoc multiple comparison test. Day × Genotype: [F_(8,180) = 2.899, p = 0.0046]; Day: [F_{(3.783,170.2)} = 17.09, p < 0.0001]; Genotype: [F_(2,45) = 46.15, p < 0.0001]. Escape latency was decreased on day 3 (⁺p = 0.0043), day 4 (⁺p = 0.0031), and day 5 (⁺p < 0.0001) in WT and day 3 (^#p = 0.0019), day 4 (^#p = 0.0173), and day 5 (^#p = 0.0036) in CPE-E342Q mice compared with day 1 respectively, but not in CPE-KO mice. n = 24 for WT, n = 12 for CPE-E342Q, n = 12 for CPE-KO. Values are mean ± SEM. b Travel distance during 5-day training of the water maze test. WT and CPE-E342Q showed no significant differences in travel distance during the 5-day training, although the CPE-KO mice tended to show a longer travel distance in days 3–5 of training compared to WT mice. Two-way repeated ANOVA analysis followed by Tukey’s post hoc multiple comparison test. Day × Genotype: [F_(8,180) = 1.702, p = 0.1007]; Day: [F_{(3.611,162.5)} = 15.40, p < 0.0001]; Genotype: [F_(2,45) = 1.148, p = 0.3265]. ⁺p = 0.0004 for WT day 5 compared with day 1; ^#p = 0.014 for CPE-E342Q day 5 compared with day1. n = 24 for WT, n = 12 for CPE-E342Q, n = 12 for CPE-KO. Values are mean ± SEM. c Time spent in each quadrant of WT, CPE-KO, and CPE-E342Q mice in Morris water maze test. WT and CPE-E342Q mice spent more time in target quadrants than CPE-KO mice which spent about the same time in each quadrant. Two-way repeated ANOVA followed by Tukey’s post hoc multiple comparison test. Area × Genotype: [F_(6,135) = 2.986, p = 0.009]; Genotype: [F_(2,45) = 14.64, p < 0.0001]; Area: [F_{(2.474,111.3)} = 8.030, p = 0.0002]. ⁺p = 0.015 for CPE-KO compared with WT mice in NE target area. SW(Southwest), NW(Northwest), NE(Northeast), SE(Southeast). n = 24 for WT, n = 12 for CPE-E342Q, n = 12 for CPE-KO. Values are mean ± SEM. d Immobility time of WT, CPE-KO, and CPE-E342Q mice in forced-swim test. Both CPE-KO and CPE-E342Q mice displayed increased immobility in the forced-swim test, suggesting depressive-like behavior. One-way ANOVA followed by Tukey’s post hoc multiple comparison test. [F_(2,45) = 15.64, p < 0.0001]. ⁺p < 0.0001 for CPE-E342Q compared with WT; ⁺p = 0.0003 for CPE-KO compared with WT. n = 24 for WT, n = 12 for CPE-E342Q, n = 12 for CPE-KO. Values are mean ± SEM. e Sucrose preference of WT, CPE-KO, and CPE-E342Q mice. CPE-KO and CPE-E342Q mice both displayed decreased preference for sucrose, suggesting depressive-like behavior. One-way ANOVA analysis followed by Tukey’s post hoc multiple comparison test. [F _(2,45) = 18.77, p < 0.0001]. ⁺p < 0.0001 for both CPE-E342Q and CPE-KO compared with WT. n = 24 for WT, n = 12 for CPE-E342Q, n = 12 for CPE-KO. Values are mean ± SEM. f Representative Western blot of CPE in WT, CPE-E342Q, and CPE-KO (which showed no expression of CPE) and (g) quantification showing CPE protein level in the hippocampus of WT, and CPE-E342Q mice subjected to chronic restraint stress. Two-way ANOVA analysis followed by Tukey’s post hoc multiple comparison test. Genotype × Restraint stress: [F_(1,12) = 0.0566, p = 0.816]; Restraint stress: [F_(1,12) = 27.81, p = 0.0002]; Genotype: [F_(1,12) = 0.5668, p = 0.4660]. ⁺p = 0.01 for restraint stressed WT compared with naïve WT; ^#p = 0.018 for restraint stressed CPE-E342Q, compared with naïve CPE-E342Q. n = 4 mice, values are mean ± SEM. h Representative western blot and (i) quantification showing BCL2 protein level in the hippocampus of WT, CPE-E342Q, and CPE-KO mice challenged to chronic restraint stress. Two-way ANOVA analysis followed by Tukey’s post hoc multiple comparison test. Genotype × Restraint stress: [F_(2,18) = 5.019, p = 0.0185]; Restraint stress: [F_(1,18) = 14.74, p = 0.0012]; Genotype: [F_(2,18) = 3.047, p = 0.0725]. ⁺p = 0.022 for restraint stressed WT compared with naïve WT; ^#p = 0.031 for restraint stressed CPE-E342Q compared with naïve CPE-E342Q. n = 4 mice, values are mean ± SEM.