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. Author manuscript; available in PMC: 2012 Oct 6.
Published in final edited form as: Neuron. 2011 Oct 6;72(1):72–85. doi: 10.1016/j.neuron.2011.08.022

Figure 7. ChIP-qPCR and immunohistochemistry confirm broad binding of total MeCP2 and global MeCP2 S421 phosphorylation in vitro and in vivo.

Figure 7

(A) ChIP-qPCR analysis of total MeCP2 (top panel) and pS421 MeCP2 (bottom panel) in cultured cortical neurons (E16 + 7 DIV) that were either membrane depolarized for 2 hours (55mM KCl) or left unstimulated.

(B) Total MeCP2 (left) and pS421 MeCP2 (right) ChIP-qPCR from the forebrains of 7-week-old MeCP2 S421A mice or their wild-type littermates. The location of PCR amplicons across the Bdnf locus are shown in diagram as a-j. Nucleotide position is given relative to start of the activity-dependent Bdnf promoter IV.

All ChIP data is presented as fold enrichment above a negative control ChIP performed in parallel on each sample using the same antiserum that had been preincubated with the peptide antigen to which it was raised. Data are mean ± SEM from three independent experiments.

(C) Immunocytochemistry of nuclei from wild-type E16 + 10 DIV mouse cortical neurons following 30 min membrane depolarization with 55 mM KCl. Antibodies used are specific for total MeCP2 or pS421 MeCP2 (see also Figure S4). Representative 63X images from single confocal planes are shown.