Figure 1.

Generation of the Isogenic Model of Rett Syndrome In Vitro

(A) Fibroblasts isolated from Rett syndrome patients (R982 and R567) heterozygous for MECP2 mutations exhibit a mosaic pattern of MECP2 expression due to random XCI.

(B) Multiple isogenic hiPSC lines were produced from patient 982 with a typical Yamanaka protocol yielding individual isogenic clones with and without MECP2 expression from the same patient, as judged by NANOG and OCT4 staining.

(C) Specification of hiPSCs derived from patient 982 toward neural progenitor cells yielded homogeneous cultures of NPCs with and without MECP2.

(D) Terminal differentiation of NPCs derived from patient 982 toward neurons and glia by growth factor withdrawal as measured by immunostaining for MAP2 and GFAP.

(E) MECP2+ and MECP2− hiPSCs and neurons were assayed by western blot with antibodies that recognize the active forms of Akt and its downstream target S6.

(F) Sholl assay of dendritic complexity was performed on WT versus MUT neurons derived from patient 982.

^∗p < 0.05 according to Student's t test. Bar graphs represent means ± SEM. Scale bars on images indicate 10 μm.