Fig. 6

Whole-mount marker studies of mouse embryos at the neurulation stage. a–f Immunohistochemical and fluorescence analyses of E8.5. Β-galactosidase (β-gal)-GRHL3 (magenta), F-actin (green), and DAPI (blue; a–d). Cre recombinase, internal ribosomal entry site (IRES), and nuclear localized β-gal were knocked in Grhl3 in the frame of the ATG start site in the Grhl3 cre allele. Arrows indicate multinucleate surface ectoderm (SE) cells (a–c) in a Grhl3^cre/+ embryo. β-gal-GRHL3 (magenta) and VANGL2 (green) in a Grhl3^cre/+ embryo (e, f). g–p VANGL2 (green; g, h), CELSR1 (green; i, j), RHOA (green; k, l), SCRIB (green; m, n), phospho non-muscle myosin light chain (pMLC green; o, p) and DAPI (blue; g–p) in wild-type (g, i, k, m, o), and Grhl3^cre/cre (h, j, l, n, p) embryos at E8.5. RHOA expression during cell division appeared to be unchanged in a Grhl3^cre/cre embryo (l). q–t Whole-mount in situ hybridization of Vangl2 mRNA in wild-type (q, s) and Grhl3^cre/cre (r, t) embryos at E8.5. u–x Molecular marker analyses in Tg(CAG-Grhl3) embryos at E3.5. Immunohistochemical analyses of RHOA (magenta; u, v), CELSR1 (magenta; w, x), and DAPI (cyan) in wild type (u, w) and Tg(CAG-Grhl3) (v, x), respectively. Representative images from more than two independent experiments. Scale bars represent 20 (u–x) and 50 μm (a–r)