Additional details can be found in the Supplemental Experimental Procedures. Total RNA was prepared Smad inhibitor from primary neurons. Additional details can be found in the Supplemental

Experimental Procedures. Primer sequences are in Table S1. Phosphatase assays were conducted by using purified calcineurin. Additional details can be found in the Supplemental Experimental Procedures. Lentiviral supernatants were prepared as described previously (Salmon and Trono, 2006). Additional details can be found in the Supplemental Experimental Procedures. Immunohistochemistry was performed on tissue sections from mouse brain. Additional details can be found in the Supplemental Experimental Procedures. Details of immunofluorescence techniques can be found in the Supplemental Experimental Procedures. Western blot scans were analyzed by using ImageJ. A rectangle was drawn around the band, and analysis was done by using the Plot Profile command. Plot Profile command displays, for a rectangular selection, a “column average plot,” in which the x axis represents the horizontal distance through the selection and the y axis indicates the vertically averaged pixel intensity. Mean values are presented with error bars corresponding to ±SEM. Statistical analysis was performed by using Prism statistical

analysis software (GraphPad). Significance ABT-888 price is indicated as ∗∗∗p < 0.001; ∗∗p < 0.01; ∗p < 0.05. A special thanks to P. Leder (Department of Genetics, Harvard Medical School) for the DAXXFlox/Flox mouse line. We also thank A. Riccio (Medical Research Council Laboratory for Molecular and Cell Biology) and G. Almouzni (Institut Curie) for discussion and critical comments on the

manuscript. We thank F. Guillemot (National Institute of Medical Research) and F. Calegari (Centre for Regenerative Montelukast Sodium Medicine) for protocols and reagents, D. Trono (School of Life Sciences and Frontiers-in-Genetics National Program, Ecole Polytechnique Fédérale de Lausanne) for providing lentiviral vectors, A. Genazzani (University of Eastern Piedmont) for ΔCAIN and calcineurin vectors, and D.L. Spector (Cold Spring Harbor Laboratory) for the YFP-H3.3/H3 plasmids. Finally, we thank D. Dinsdale and J. Edwards (Medical Research Council Toxicology Unit) for support and assistance with histology and S. Beck (University College London Cancer Institute) and C. Widmann (Institute of Physiology, University of Lausanne) for critical discussion. P.S., D.M. and S.B. are supported by the Samantha Dickson Brain Cancer Trust. P.S. is also funded by the Wellcome Trust. “
“Synapses need to be functionally and structurally maintained throughout life to preserve stable neuronal networks and normal behavior (Holtmaat and Svoboda, 2009 and Lin and Koleske, 2010). Longitudinal in vivo imaging in mice has shown that the majority of synapses are stable for a lifetime (Grutzendler and Gan, 2006 and Holtmaat et al., 2006).