Mice were anesthetized and then perfused, cryopreserved, embedded, and sectioned as previously described (Liu et al., 2012 (link)). Immunohistochemistry was performed as previously described (Liu et al., 2012 (link)) with primary antibodies against trimethylated histone 3 lysine 9 (H3K9me3, 1:100; ab8898, Abcam), CC1 (1:100; OP80, Calbiochem), myelin basic protein (MBP, 1:500; SMI99, Covance), OLIG2 (1:200, ab81093, Abcam), NG2 (1:200; AB5320, EMD Millipore) or Caspr (1:100, ab34151, Abcam). Stained sections were visualized using confocal microscopy (LSM800 Meta confocal laser scanning microscope, Carl Zeiss Micro-Imaging). For NG2, CC1, OLIG2 cell counts, and H3K9me3 intensity quantifications, 4–6 20x fields were taken per mouse. For MBP-covered segments and internodal length marked by Caspr, 4–6 fields were taken per mouse followed by quantifications using ImageJ. One-way ANOVA tests were performed to assess statistical differences. For correlation of internodal length with social interaction ratio, data normality was determined using D’Agostino and Person test in GraphPad Prism 8. Pearson correlation coefficients were calculated if data passed normality test.
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