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. Author manuscript; available in PMC: 2014 Mar 21.
Published in final edited form as: Neuron. 2006 Oct 19;52(2):255–269. doi: 10.1016/j.neuron.2006.09.037

Figure 6. Phosphorylation of MeCP2 at S421 Mediates MeCP2-Dependent Regulation of Dendritic Growth and Spine Maturation.

Figure 6

(A) Organotypic hippocampal slices were prepared from P5–7 rat pups and biolistically transfected after 2 days in culture with LEMPRA-based plasmid DNA encoding a control vector (in which the shRNA directed against MeCP2 is replaced with a scrambled shRNA and a stop codon is introduced directly after the FLAG epitope), the anti-MeCP2 shRNA only, wild-type MeCP2, or S421A MeCP2, in combination with a dual-promoter plasmid expressing eGFP and Bcl-XL. Representative 25× images of 7 DIV transfected pyramidal neurons are shown. Scale bar, 50 μm. Pseudocoloring: green, GFP; red, anti-FLAG; blue, antitotal MeCP2.

(B) Representative 63× GFP images of dendritic spines from control-, anti-MeCP2 shRNA-, wild-type MeCP2-, or S421A MeCP2-transfected neurons. Scale bar, 2 μm.

(C and D) Quantification of dendritic branch complexity by Sholl analysis (data are presented as mean ± SEM, *p < 0.05, ANOVA).

(E and F) shRNA-mediated knockdown of MeCP2 does not affect dendritic spine morphology (p > 0.05, K-S test)

(G and H) Quantification of the effects of wild-type or S421A mutant MeCP2 overexpression on dendritic spine length (G) and width (H) (p < 0.05 for wild-type versus S421A or wild-type versus control, K-S test).