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. 2013 Dec 18;23(9):2447–2458. doi: 10.1093/hmg/ddt640

Figure 5.

Figure 5.

Western blot analysis of brain MeCP2 isoform levels confirms loss of MeCP2-e1 and overexpression of MeCP2-e2. Total brain proteins from Mecp2-e1−/y and Mecp2-e1+/y control males were probed with an MeCP2-e1 CS specific antibody in (A) which shows barely detectable MeCP2-e1 levels in Mecp2-e1−/y brain and significantly higher levels in WT control male brain (P = 0.0019). (B) An anti-MeCP2-e2-specific antibody revealed significant elevation of this isoform (P = 0.0005) in Mecp2-e1−/y brain compared with WT (Mecp2-e1+/y) controls. (C) To determine the combined levels of MeCP2-e1 and MeCP2-e2 isoforms, an antibody recognizing a common epitope (Fig. S1) revealed an ∼2-fold reduction of total MeCP2 in Mecp2-e1−/y deficient brain by western analysis (P = 0.01). Bar graphs depict average signal intensity from NIH Image J analysis of western blots with error bars corresponding to SEM. Three to four individual mouse brain protein extracts per genotype were analyzed.