Fig. 4: BTBR/R showed behavioral deficits and autistic phenotypes differently than BTBR/J.

All the behavioral tests were analyzed in both BTBR strains and compared to B6. a Open field test in a 30-min section. The level of anxiety was accessed by the percentage of time spent in the center region. B6, n = 17; BTBR/R, n = 22; BTBR/J, n = 14. One-way ANOVA (effect of genotype), F_2,50 = 22.89, P < 0.0001. b Light-dark box for 10 min. The level of anxiety was accessed by the time spent in the dark box. B6, n = 18; BTBR/R, n = 17; BTBR/J, n = 8. One-way ANOVA (effect of genotype), F_2,40 = 13.43, P < 0.0001. c Pup USV calls. Total call number emitted by P8 pups within 3 min of maternal separation. B6, n = 17; BTBR/R, n = 26; BTBR/J, n = 28. One-way ANOVA (effect of genotype), F_2,68 = 6.330, P = 0.0030. d Different preferences for call types of multiple syllables between two BTBR strains. Ts, two-syllable; Fs, frequency steps, Cp, composite; Har, harmonics. Unpaired t-test. e Call type usage summarized in pie charts for each strain. USV calls with single syllables were classified as “simple”. Calls of two-syllable and frequency steps were grouped as “sequential”; Calls of composite and harmonics were grouped as “overlapping” in the pie chart. f Adult male to female courtship calls within 5 min session. B6, n = 15; BTBR/R, n = 22; BTBR/J, n = 12. One-way ANOVA (effect of genotype), F_2,46 = 21.89, P < 0.0001. g Repetitive behaviors analyzed by self-grooming in 10 min. B6, n = 14; BTBR/R, n = 22; BTBR/J, n = 14. One-way ANOVA (effect of genotype), F_2,47 = 22.62, P < 0.0001. h Repetitive behaviors analyzed by marble burying in a 30 min section. The statistic result for the number of buried marbles. B6, n = 18; BTBR/R, n = 18; BTBR/J, n = 13. Still, one-way ANOVA (effect of genotype), F_2,46 = 11.78, P < 0.0001; buried, one-way ANOVA (effect of genotype), F_2,46 = 12.34, P < 0.0001; half, one-way ANOVA (effect of genotype), F_2,46 = 5.208, P = 0.0091; moved, one-way ANOVA (effect of genotype), F_2,46 = 0.3854, P = 0.6824. i Social interaction analyzed by three-chambered apparatus. The first 10-min section tested the sociability of the subject by analyzing its preference between a stranger mouse (stranger 1) and an empty cage. The second 10-min section tested the preference for social novelty by accessing the time spent on stranger 1 (more familiar) and stranger 2 mouse (novel). B6, n = 11; BTBR/R, n = 16; BTBR/J, n = 11. Unpaired t-test. j Spatial learning memory measured by Barnes maze. Left, the spatial performance profile of the 6-day training. B6, n = 18; BTBR/R, n = 21; BTBR/J, n = 13. D4, one-way ANOVA (effect of genotype), F_2,49 = 5.983, P = 0.0047. D6, one-way ANOVA (effect of genotype), F_2,49 = 6.782, P = 0.0025. Right, probe test on day 7. One-way ANOVA (effect of genotype), F_2,49 = 7.390, P = 0.0016. B6, n = 11; BTBR/R, n = 16; BTBR/J, n = 11. Unpaired t-test. Data shown are mean (±S.E.M.) for each strain and analyzed by one-way ANOVA followed by Tukey’s multiple comparison test, ***, p < 0.001; **, p < 0.01; *, p < 0.05; n.s., not significant).