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Fig. 2 | Epigenetics & Chromatin

Fig. 2

From: MeCP2-E1 isoform is a dynamically expressed, weakly DNA-bound protein with different protein and DNA interactions compared to MeCP2-E2

Fig. 2

Isoforms N-terminal processing, turn-over rates, and dynamics. a Mass spectrometry sequencing of the N-terminal end of the MeCP2 protein (in vitro). N-terminal peptide coverage alignment chart and high-resolution mass spectra showing N-methionine excision (NME) and N-acetylation (NA) of the N-termini of MeCP2-E1 and MeCP2-E2. NA (+42 Da) of N-terminus amino acid is shown highlighted in yellow. bd Cycloheximide-chase assays of the E1 and E2 MeCP2 isoforms. Densitometries and representative western blots performed after cycloheximide treatments of b SH-SY5Y cells overexpressing (OE) E1 and E2 isoforms fused to GFP, c differentiated SH-SY5Y, and d rat cultured cortical neurons with detection of endogenous E1 and E2 isoforms. e Densitometric analysis and representative western blots showing endogenous E1 and E2 levels in frontal cortices of mice euthanized at 12 a.m. and 12 p.m. f KCL depolarization and representative Western blots analysis of total endogenous MeCP2 of cultured cortical neurons and E1 and E2 of transfected cultured cortical neurons overexpressing Flag-MeCP2-E1 or E2. Represented data are mean ± S.E.M. (n = 7–8). * P < 0.05 two-tailed Mann–Whitney test. MeCP2 levels were normalized using β actin and/or histone H3

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