Fig. 2.

Behavioral analysis of Cyld^−/−, Shank3^−/−, and Cyld^−/−Shank3^−/− double-mutant mice. (A and B) The vocalization analysis of the number of calls emitted in the presence of a female mouse and latency to first call. (C and D) The repetitive behavior assessment with both marble burying test and time of grooming. (E) The analysis of the exploratory behavior with the time of the first and the second exposure to the same object. (F–I) The analysis of spatial memory by MWM test with measures of total distance traveled and latency to the platform during the hidden training phase and reverse phase. For experiments in A and B, n = 15 for Cyld^+/+Shank3^+/+ controls (one excluded, no vocalization), n = 22 for Cyld^−/− mice (four excluded, no vocalization), n = 17 for Shank3^−/− (one excluded, no vocalization), and n = 14 for Cyld^−/−Shank3^−/− mice (three excluded, no vocalization) from four independent cohorts. For experiments in C through E, n = 16 for Cyld^+/+Shank3^+/+ controls, n = 26 for Cyld^−/− mice, n = 18 for Shank3^−/−, and n = 17 for Cyld^−/−Shank3^−/− mice from four independent cohorts. For experiments in F through I, n = 15 for Cyld^+/+Shank3^+/+ controls, n = 22 for Cyld^−/− mice, n = 17 for Shank3^−/−, and n = 13 for Cyld^−/−Shank3^−/− mice from four independent cohorts. All n values used for statistics refer to the number of mice used after identification of possible outliers with Grubbs' method. Statistics were calculated by ordinary one-way ANOVA followed by Tukey’s multiple comparisons test (A–D) by unpaired nonparametric Mann–Whitney U test (E) or by two-way repeated-measures ANOVA with Dunnett’s multiple comparisons test (F–H). *P < 0.05, **P < 0.01, graphs are mean ± SEM.