Fig. S1. Tissue disorganization and loss of the hippocampus in the brains of β-catenin conditional knockout (KO) embryos. (A) Filamentous actin (F-actin), as revealed by staining with rhodamine-phalloidin, is normally distributed in cells lining the lateral ventricle (LV) (arrows) at E14.5. Blue, 4′,6-diamidino-2-phenylindole (DAPI) staining of DNA. (B) In conditional KO embryos, F-actin accumulation is prevalent in deeper regions of the striatum (STR) (arrows). (C) Normally, N-cadherin (Ncad) occupies the lining of the LV (arrows). (D) In conditional KO brains, N-cadherin protein is more highly concentrated in apical regions (arrows). (E) Nestin is expressed in the periventricular region of control STR. (F) In conditional KO STR, nestin is still expressed in the same region despite significant tissue disorganization. Mash1 is similarly expressed in the periventricular region of both the control (G) and KO (H) STR. CTX, cortex.

Fig. S2. (A) In E17 wildtype cortex (CTX), Tbr1 staining (red) delineates the cortical plate (CP) and intermediate zone (IZ), whereas Pax6 staining (green) is restricted to the ventricular zone (VZ). Note that some of the Pax6 staining in the IZ may be attributed to background binding of the primary antibody to mouse IgG. (B) A more specific rabbit polyclonal antibody to Pax6 demarcates the VZ of the wildtype CTX. (C) In Nestin-mediated β-catenin knockout (KO) embryos, Tbr1 expression extends ventrally toward the ventricle resulting in overlap of markers of the intermediate and ventricular cortical zone cells. (D) Extension of Pax6-positive VZ cells into more dorsal regions of KO CTX. (E) Prox1 (red) is expressed in the hippocampal primordium of E17 wildtype embryos. HC, hippocampus. (F) Higher magnification view of box in E showing Prox1-positive nuclei. (G) The neuroepithelium of mutant cortices is thinner. Also, much of the HC is lost in mutant mice. (H) Higher magnification view of box in E showing reduced number of Prox1-positive nuclei.

Fig. S3. Genotyping of neurosphere cultures to detect loxed (lox), wildtype (wt) and deleted (del) β-catenin alleles. Lane 1, Knockout (ko) culture that is Nestin:Cre positive and therefore does not possess a loxed or a wildtype β-catenin allele. Lane 2, Wildtype culture that does not carry the Nestin:Cre allele and that has one loxed β-catenin allele and one wildtype allele. Lane 3, Knockout culture that has a deleted β-catenin allele. Lane 4, Wildtype culture that does not have a deleted β-catenin allele. PCR, polymerase chain reaction.

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