A detailed description of animal breeding and genotyping is provided in the Supplemental Experimental Procedures. Cortical or hippocampal neurons were prepared from E16 mouse embryos as described by Kim et al. (2010). Western blotting, immunocytochemistry,

and immunohistochemistry were carried out using standard methods. Sholl analysis of dendritic morphology was performed using Neurolucida (MBF Bioscience) software Galunisertib on images acquired by confocal microscopy. For details see Supplemental Experimental Procedures. ChIP experiments were performed as previously described (Kim et al., 2010) with some modifications. Immunoprecipitation was performed using custom rabbit polyclonal antisera recognizing either total MeCP2 or phosphoserine 421 MeCP2 (Zhou et al., 2006). ChIP-Seq was performed using the SOLiD instrument (Life Technologies) as previously described (Kim et al., 2010), with slight modifications. For a description of computational analysis and experimental details see Supplemental Experimental Procedures. For gene expression in cultured neurons or brain tissue, total RNA was isolated and cDNA generated using standard methods. qPCR analysis was carried out using the StepOnePlus system (Life Technologies, Beverly, MA). Microarray analysis of cultured wild-type and S421A neuron

RNA samples was carried out using Affymetrix GeneChip Tenofovir mw Mouse Expression Set 430 2.0 microarrays arrays. Microarray analysis from visual cortex employed total RNA isolated from P17 mice and analyzed on Affymetrix Mouse Gene 1.0 ST microarrays. See Supplemental Experimental Procedures for details. Whole-cell voltage clamp recordings were measured mafosfamide from layer II/III pyramidal neurons in coronal sections (250 μm) containing the primary visual cortex of P16–P17 mice. For both amplitude and interevent interval, 250 events

were randomly selected from each cell. See Supplemental Experimental Procedures for details. The assay of sociability and preference for social novelty were based on a well-characterized assay from the Crawley lab (Moy et al., 2004). For details of all behavioral analyses see Supplemental Experimental Procedures. We thank E. Griffith, P. Greer, S. Ross, N. Robinson, and Greenberg Lab members for critical reading of the manuscript; C. Chen, B. Bloodgood, E. Hong, R. Rodriguiz, J. Dodart, H. Ho, and the MRDDRC Gene Manipulation Core (M. Thompson, Y. Zhou, and H. Ye) for experimental assistance and advice. This work was supported by NIH grant 1R01NS048276 to M.E.G. A.E.W. was supported by NIH grant R01 DA022202 and March of Dimes grant 1-FY07-482; S.C. was supported by the Medical Scientist Training Program, Ruth L. Kirschstein NIH Training Grant T32CA009361, and the Department of Neurobiology Graduate Fellowship Fund at HMS; H.W.G. was supported by Damon Runyon Cancer Research Foundation Grant DRG-2048-10; D.H.E.