Fig. S3

(A-I) her2 is partially dependent on Notch signaling and id2a, id3 and id4, unlike id1, are decreased in mecp2 morphants. (A-B′) her2 expression was slightly up-regulated in embryos injected with NICD-mRNA (75 pg). (C) Quantitative real-time PCR analysis of her2 expression in wild-type embryos at 24 hpf and 48 hpf, normalized to gapdh. Measurements are the means from 3 independent experiments ± SD. *P <0.05. (D-D′) her2 was slightly down-regulated in mecp2 morphants treated with DAPT (100 µM). (E) Quantitative real-time PCR analysis of her2 expression in mecp2 morphants with and without DAPT, normalized to gapdh. The data represent the means of three independent experiments ± SD. *P <0.05. (F-H′) RNA in situ hybridization with id2a, id3, and id4 probes in controls (F, G, H ) and mecp2 morphants (F′, G′, H′) at 48 hpf. Note that these genes decreased in the developing brain of mecp2 morphants. Dorsal views are shown with anterior to the left (A-B′, D-D′, F-H′). The numbers in the bottom right corner represent phenotypic embryos/total embryos. (I) Quantitative real-time PCR analysis of id2a, id3, and id4 in wild-type and mecp2 morphant embryos at 48 hpf, normalized to gapdh. Measurements are the means from 3 independent experiments ± SD. *P <0.05. (J-M) Both id1-MO and mecp2-mRNA rescue the phenotype in mecp2 morphants. RNA in situ hybridization with nestin, gfap, map2 and neurod probes in controls (J-M), mecp2 morphants (J′-M′), mecp2 morphants co-injected with id1-MO (J′′-M′′) and mecp2 morphants co-injected with mecp2-mRNA (J′′′-M′′′) at 48 hpf. Note that expression of these genes in the developing brain in mecp2 morphants was rescued by co-injection of either id1-MO or mecp2-mRNA. Dorsal views are shown with anterior to the left. The numbers in the bottom right corner represent phenotypic embryos/total embryos.