Fig. 3
Grhl2 regulates a molecular signature of epithelia in the surface ectoderm. a Schematic representation of an E9.5 embryo and caudal spinal region isolated for analysis. b–c qRT-PCR of Cdh1, EpCAM, Cldn 3,4,6,7, and 8 of samples derived from the whole-caudal spinal region (as in a) (n = 3–6 samples per genotype). d Schematic representation of the microdissection separating the dorsal (surface-ectoderm-containing) and the ventral (hindgut-containing) parts of the caudal spinal region. e, f qRT-PCR analysis of Cdh1, EpCAM, Cldn 3,4,6,7, and 8 of samples derived from microdissected spinal regions (as in d) ( > 5 embryos pooled per sample; three samples per genotype). Data represent mean ± SEM (****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05; t-test); (See Supplementary Fig. 3 for equivalent Grhl2−/− data). WMISH confirmed upregulation of Cdh1 (g–l), Cldn4 (m–r), and EpCAM (s–x) in Axd/Axd compared with +/+ embryos at E9.5 (examples at 17–19 somite stage). Transverse sections are at the levels indicated by dashed lines in g (se, surface ectoderm; hg, hindgut). Scale bars represent 100 µm in whole mounts and 25 µm on sections. y–dʹ Dorsal view of PNP region (caudal end oriented upwards; asterisk indicates closure site) immunostained for E-cadherin (y, z) and Cldn4 (aʹ–dʹ). bʹ and dʹ show magnified images of areas boxed in a' and c', respectively. Scale bars indicate 25 μm in y, z, aʹ, cʹ and 10 μm in bʹ and dʹ. Images are representative of a minimum of three embryos per genotype. See also Supplementary Fig. 4 for additional WMISH of Cldn genes in Axd and Supplementary Fig. 5 for equivalent data in Grhl2−/−. Source data are provided as a Source Data file
