Figure 2

Mecp2 mRNA and protein are present in the brains of A140V mutant male mice. (2A) Total RNA was extracted from the cerebella of mice and used for cDNA synthesis. RT-PCR confirmed the presence of Mecp2 mRNA in the cerebella of 2 WT and 2 A140V (Mut) mice (Left Panel). The mutant mRNA species results in a larger fragment by PCR than the WT mRNA species. A second RT-PCR reaction amplifying a different segment of the cDNA containing the A140V mutation site confirmed that there was no contamination of the total RNA used for the cDNA synthesis with genomic DNA. WT and A140V (Mut) cDNA showed only the presence of a band of the correct size for cDNA PCR product (Right Panel). (2B) Immunohistochemistry on frozen brain slices demonstrates the presence of MeCP2 protein in the brain of an A140V hemizygous mouse. Staining in the hippocampus and the cerebellum are similar in the WT and A140V mice using an antibody recognizing the C-terminus of MeCP2. Scale bars are 200 μm.