Figure 1. Morphological defects in the Pten mutant cephalic neural plate.
(A, B, C, D) Comparison of neural plate morphology of the dorsal head of wild-type (WT) and Pten △Epi mutant embryos at E8.0 and E8.5 in scanning electron microscope images. Scale bar = 100 μm. (E, F) Transverse sections of E8.5 WT and Pten △Epi embryos show the absence of pseudostratified columnar organization in the Pten mutant cephalic neural plate. Green is SOX2, red is phalloidin (F-actin), blue is DAPI. (G, H) Z-stack projection of three optical sections (total of 3 µm) from transverse sections of the cephalic neural plate of E8.5 WT and Pten △Epi mutant embryos stained for phalloidin (red) and laminin (purple). Scale bar E–H = 10 μm.
Figure 1—figure supplement 1. PTEN expression in the cephalic neural plate.
Immunodetection of PTEN in transverse sections of E8.5 WT (A, C, E) and the Pten △Epi embryos (B, D, F) shows the specificity of PTEN staining. (A,B) Antibody from cell signaling; (C–F) antibody from cascade. (G, H) Z-stack projections of 3 optical sections taken every 1 μm, showing expression pAKT S473 expression in transverse sections of the E8.5 WT (G) and Pten △Epi (H) neural plate. Red is phalloidin and blue is DAPI. Scale bar in A–D = 100 μm; in E–H = 10 μm
Figure 1—figure supplement 2. Normal neural patterning in Pten△Epi embryos.
(A) In situ hybridization for Engrailed2, Krox20, Emx2 and Fgf8 in WT and Pten △Epi embryos at E8.5 showed normal anteroposterior regionalization of cephalic neural epithelium. Scale bar = 180 μm. (B) Immunodetection of FoxA2, Nkx2.2 and Nkx6.1 shows normal dorsal-ventral patterning of the E8.5 cephalic neural plate in Pten △Epi embryos. Scale bar = 100 μm. (C) The TOPGAL reporter of canonical Wnt signaling was expressed in the normal domains and at normal levels in E8.5 Pten △Epi embryos. Scale bar = 180 μm.
Figure 1—figure supplement 3. Apical markers in Pten △Epi mutant embryos.
Z-stack projection of three optical sections (total of 3 µm) from transverse sections of the cephalic neural plate of E8.5 WT and Pten △Epi mutant embryos stained for (A, B) N-Cadherin, (C, D) pERM, (E, F) ZO1 and (G, H) aPKC (green) and PAR3 (red). (I) Comparison of the number of rows of nuclei in the pseudostratified cephalic neural plate of E8.5 wild-type and Pten △Epi embryos. n = 27 embryos for each genotype. Blue is DAPI. Scale bar = 20 µm.