Fig. 2
Transplantation of the fecal microbiota from EphB6-deficient mice caused autism-like behavior in 3-week-old SPF C57BL/6J mice. a–i Schematic of the fecal microbiota transplantation (a). The 3-week-old SPF male C57BL/6J mice were orally gavaged with the fecal microbiota from 8-week-old male WT or KO mice (each contained eight healthy mice from at least three cages) for 1 week. After 3 weeks, the fecal microbiota of the treated C57BL/6J mice were sequenced (b–d, eight treated C57BL/6J mice of each group were selected randomly from at least three cages) and self-grooming test (e), olfactory habituation/dishabituation test (f), three-chambered social approach task (g–i), open field test, and elevated plus maze test were conducted with an interval of at least 2 days (e–i, n = 13, 19 mice respectively). j–r Schematic of the fecal microbiota transplantation (j). The 3-week-old SPF male C57BL/6J mice were orally gavaged with antibiotics (ampicillin, vancomycin, neomycin, metronidazole) for 5 days and then orally gavaged with the fecal microbiota from 8-week-old male WT or KO mice (each contained eight healthy mice from at least three cages) for another 5 days. After 19 days, the fecal microbiota of the treated C57BL/6J mice were sequenced (k–m, six treated C57BL/6J mice of each group were selected randomly from at least two cages) and self-grooming test (n), olfactory habituation/dishabituation test (o), three-chambered social approach task (p–r), open field test, and elevated plus maze test were conducted with an interval of at least 2 days (n–r, n = 10, 16 mice respectively). Data shown are mean ± SEM or median ± IQR. Two-tailed unpaired student’s t test (e, h, i, n, q, r), Mann-Whitney test (b, c, k, l), mixed design ANOVA with genotype as independent factor and stimuli/trials as repeated-measure factor (f, o), anosim analysis (d, m). *p < 0.05. WT col or KO col colonized with the fecal microbiota from EphB6+/+ or EphB6−/− mice, Abx pretreated with antibiotics (ampicillin, vancomycin, neomycin, metronidazole). Statistical values are presented in Additional file 3: Table S2