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. 2024 Jan 26;25(3):1543. doi: 10.3390/ijms25031543

Figure 4.

Figure 4

Analysis of social behavior in wild-type, heterozygous, and homozygous Uhrf2 knockout mice using the three-chamber sociability and social novelty test. (A) The figure illustrates a schematic representation of the three-chamber social test. (B) The total distance traveled by mice with different genotypes was measured during the habituation period. A two-way RM ANOVA revealed a significant effect of genotype (F (2, 18) = 6.33, ** p = 0.008), but no significant effect of test day (F (2, 36) = 0.21, p = 0.82, ns). On the first day (D1), there was no significant difference in distance traveled among the three genotypes: wild-type mice (Uhrf2+/+, gray, n = 8), heterozygous mice (Uhrf2+/−, green, n = 6), and knockout mice (Uhrf2−/−, red, n = 7). However, on day 2 (D2), the heterozygous mice moved significantly shorter distances compared to the knockout mice (* p = 0.017). On day 3 (D3), the homozygous knockout mice traveled significantly longer distances than both the wild-type and heterozygous mice (** p = 0.001, * p = 0.02). (C) The velocity of mice with different genotypes was measured during the habituation period. Statistical analysis using a two-way RM ANOVA revealed a significant effect of genotype (F (2, 18) = 7.45, p = 0.004), while no significant effect of test days was observed (F (2, 36) = 1.59, p = 0.21; ns). On days 2 and 3, knockout mice traveled at a significantly higher velocity than wild-type mice (* p = 0.036, *** p < 0.0003). (D) The exploration preference of the mice during the habituation period on the test day (D4) was evaluated. A two-way RM ANOVA showed no significant effect of genotype (F (2, 18) = 3.37, p = 0.06, ns) and no significant effect of chamber (F (1, 18) = 0.01, ns) on the outcome of the test. There were no significant differences in exploration time between wild-type, heterozygous, and knockout mice in the left and right chambers (ns, not significant). (E) Sociability was assessed by measuring the amount of time mice spent exploring the familiar mouse compared to the empty cage. Wild-type mice spent significantly more time exploring the familiar mouse than the empty cage (* p = 0.043). However, no significant difference was observed between the exploration of the familiar mouse and the empty chamber in heterozygous and knockout mice (ns, not significant). A two-way repeated measures ANOVA revealed that the effect of genotype was not significant (F (2, 18) = 2.07, p = 0.16, ns), but the effect of chamber was significant (F (1, 18) = 5.16, p = 0.0357). (F) In terms of social novelty in mice, wild-type mice exhibited significantly higher levels of exploration towards the unfamiliar mouse compared to the familiar mouse (** p = 0.0025). Furthermore, no significant difference in exploration time between the unfamiliar and familiar mice was observed in heterozygous and knockout mice. A two-way repeated measures ANOVA revealed an effect of the chamber on the variations (F (1, 18) = 11.68, p = 0.003), but no significant effect of genotype (F (2, 18) = 0.94, p = 0.41). The p-values presented in this legend represent the results of comparisons analyzed using Sidak’s multiple comparisons.