Mecp2tm1.1bird j

Mecp2^tm1.1Bird/J and control female mice were purchased from The Jackson Laboratories. Genotyping of MeCP2^+/− was performed by PCR analysis of tail DNA (9875: AAA TTG GGT TAC ACC GCT GA, Common; 9877: CCA CCT AGC CTG CCT GTA CT, Mutant Reverse; 7172: CTG TAT CCT TGG GTC AAG CTG, WT Reverse). All experiments were conducted according to the ARRIVE guidelines (Animal Research: Reporting In Vivo Experiments). Animals were randomized and coded so that the persons carrying out behavioural analysis, tissue processing and statistical analysis were blind to the treatment. Randomization was carried out using the Research Randomizer Program online (www.randomizer.org). The GPower program was used to calculate a priori the sample size. Power, alpha and effect size were set at 80%, 0.05 and 0.44, respectively. The number of subjects in the experiments is reported in the figure legends. All experiments with mice were performed according to the national and international laws for laboratory animal welfare and experimentation (EU directive n. 2010/63/EU and Italian DL n. 26 04/03/2014). Mice were kept under a 12-h dark to light cycle, with food and water ad libitum.

Female Mecp2 heterozygous females [54 (link)] were purchased from Jackson Laboratories (strain name: B6.129P2(C) − Mecp2^tm1.1Bird/J, stock number 003890). Mecp2 mutant hemizygous males (Mecp2 -/Y, also indicated as Mecp2 null) and wild type (WT) littermates used as controls were obtained by mating heterozygous females with WT males (C57BL6/J background) for at least 12 generations. The national or institutional guidelines were used for the care and use of animals, and approval for the experiments were obtained from the ethical committees of the Italian Ministry of Health and the UK Home Office.
Genomic DNA was extracted from tail tips of embryos and immediately genotyped by polymerase chain reaction to determine Mecp2 deletion (according to [54 (link)]) and mice gender. The gender was determined using primers mSRY.F 5′-CCCAGCAGAATCCCAGCATGC-3′ and mSRYl.R 5′-TCCTGTCCCACTGCAGAAGGT-3′ specific for the Y chromosome-linked Sry gene.
Cortices from each embryo were separately dissected from those of the other littermates.