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Additional Supporting Information may be found in the online version of this article.

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SC-11-1213_sm_SupplFigure1.tif3066KSup. Fig. 1 BDNF addition does not rescue the small nucleus phenotype BDNF (40 ng/ml) was added after 2 DIV and every other day thereafter for a total period of 10 DIV. No changes were observed in nuclear size (monitored here with Lamin B antibodies) compared with untreated wild-type or mutant neurons (stained with MAP2 antibodies). ESCD refers to ES cell-derived neurons. n.s.: not significant.
SC-11-1213_sm_SupplFigure2.tif118KSup. Fig. 2. Quantification of 2 MeCP2 splice variants in cultured astrocytes and ES cell-derived neurons Both MeCP2 splice variants were quantified by qPCR using RNA extracted from cultured astrocytes (14 DIV) and ES cell-derived neurons (ESCD neurons, 14 DIV). E1 refers to the longer and E2 to the shorter MeCP2 transcript16. n.s.: not significant.
SC-11-1213_sm_SupplFigure3.pdf446KSup. Fig. 3 Co-cultures of ES cell-derived neurons with wild-type or mutant astrocytes Mecp2+/y or Mecp2−/y neurons were grown between 2 and 10 DIV in the presence of Mecp2+/y or Mecp2−/y cortical astrocytes. Nuclei were visualized with Hoechst and Lamin B staining while neuronal nuclei were identified on the basis of MAP2 staining. Note that also under these co-culture conditions it is the genotype of the neurons that determines the size of the nucleus, irrespective of the genotype of the astrocytes. ***P<0.0001, Student's t test, N=86 nuclei per condition. n.s.: not significant.

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