NexCre cTKO animals show an autism spectrum disorder ASD-like phenotype with repetitive behavior and impairments in social communication and interactionA: Diurnal behavior in the home cage (HC).Figure 5A: Diurnal behavior in the home cage (HC). NexCre cTKOs show overshooting activity in the dark phase. Line graphs illustrate the average locomotion per hour. [Sidak's multiple comparisons test, geno F(1,32) = 9.503, p = 0.0042; timepoint F(6.102,195.3) = 10.51, p < 0.0001; timepoint × geno F(47,1504) = 5.874, p < 0.0001]. B: HC, climbing event are elevated in NexCre cTKO animals during the dark phase. During the light phase NexCre cTKOs climb significantly less than LM controls. [Sidak's multiple comparisons test, geno F(1,32) = 6.218, p = 0.0180; phase F(1,32) = 29.16, p < 0.0001 ; phase × geno F(1,32) = 13.88, p = 0.0008]. C: HC, Rearing events are elevated in NexCre cTKO animals during the dark phase. [Sidak's multiple comparisons test, geno F(1,32) = 6.565, p = 0.0153; phase F(1,32) = 133.5, p < 0.0001 ; phase × geno F(1,32) = 20.05, p < 0.0001]. D: Ultrasonic Vocalisation (USV), NexCre cTKOs show a strong reduction of emitted calls compared to LM controls. [unpaired Student's t-test ***p = 0.0005]. E: USV, Compared to LM controls, the duration of time spent with call emission is strongly reduced in NexCre cTKOs. [Mann-Whitney test ****p < 0.0001]. F: USV, NexCre cTKO animals repeat calls of a certain type more frequently. [Welch's t-test *p = 0.0131] G: USV, Analysis of call type complexity. NexCre cTKOs used simple calls either with similar or reduced frequency (downwards). Note that NexCre cTKOs used complex call types less frequently. Simple call types: Short (Mann Whitney test; ns, p = 0.0747), Flat (Mann Whitney test; ns, p = 0.2172), Upwards (Mann Whitney test; ns, p = 0.2383), Downwards (Unpaired Student's t-test; LM n = 1359 calls, NexCre cTKO n = 1507 calls; **p = 0.0085) and Chevre (Welch's test; ns, p = 0.1281). Complex call types are Two Syllable (Mann Whitney test; ns, p = 0.2380), Frequency-Steps (Mann Whitney test; ns, p = 0.5246), Composite (Unpaired Student's t-test; LM = 2258 calls, NexCre cTKO n = 722 calls; **p = 0.0086), Waves (Welch's test; LM = 403 calls, NexCre cTKO n = 88 calls; **p = 0.0039) and Harmonics (Mann Whitney test; LM = 711 calls, NexCre cTKO n = 197 calls; ****p < 0.0001). H: USV, mean call type complexity. The ratio of complex calls is reduced in NexCre cTKOs in favor of simple call type emission. [two-way ANOVA, complexity F(1,27) = 44.11, p < 0.0001 ; complexity × geno F(1,27) = 9.751, p = 0.0042]. I: Sociability in the three chambers test (3CT). LM controls spend more time with mouse stranger 1 (S1) whereas NexCre cTKOs spend comparable time exploring the novel object (NO) or S1. Bottom: schematic representation of test. [Paired Student's t-test comparison between chambers LM ***p = 0.0002; Wilcoxon test between chambers NexCre cTKO ns p = 0.5412]. J: 3CT, test for social novelty. Unlike LM controls, NexCre cTKO do not spend more time with an unfamiliar stranger mouse S2 compared to S1. [Paired Student's t-test between chambers LM ****p < 0.0001; Wilcoxon test between chambers NexCre cTKO ns p = 0.1084].