a, Breeding strategy to generate mice that have a wild-type MECP2 allele and a Mecp2 allele flanked by loxP sequences. b, Western blot of MeCP2 in hypothalamus, amygdala and cerebellum. GAPDH was used as the internal control (for gel source data, see Supplementary Fig. 3). c, Densitometric analysis of western blots in b. Flox;TG mice overexpress MeCP2 at levels similar to transgenic mice. It is noteworthy that in the hypothalamus, Flox mice were 30% hypomorphic (n = 2 mice per group; two-tailed t-test) when compared to wild-type mice, but not in the other regions. d, RT–qPCR analysis in hypothalamus, amygdala and cerebellum, using primers common to mouse and human MECP2. Flox;TG mice overexpressed the MECP2 transcript at levels similar to transgenic mice. Hprt1 was used as the internal control (n = 5 mice per group; two-tailed t-test). e, RT–qPCR analysis of three selected genes know to be altered by MeCP2 overexpression. Flox;TG mice overexpressed the Sst, Crf and Prl2c2 transcripts at levels similar to those of transgenic mice. Hprt1 was used as the internal control (n = 4 for wild-type group; n = 6 for transgenic group; n = 5 for Flox and Flox;TG groups; two-tailed t-test). Data are mean ± s.e.m. *P < 0.05; **P < 0.01; ***P < 0.001.