Fig. 2

Transcriptomic effect of Grhl2 loss and over-expression. Abundance of a, b Grhl2 mRNA and c, d Grhl2 protein is significantly elevated in the caudal region of a, c Axd/Axd embryos and diminished in b, d Grhl2^−/− embryos at E9.5 (17–19 somite stage); ****p < 0.0001; t-test; mRNA data represents mean ± SEM (n = 6 samples/genotype). Example immunoblot in d is exposed to show all +/+ bands. e–k Increased expression of Grhl2 in Axd/Axd embryos at E8.5 (e, f; dorsal view in f and h) and E9.5 (i–k; examples at 16 somite stage). Apparent upregulation or ectopic expression is detected in the tail bud (tb; i–k) in Axd/Axd embryos. Scale bars represent 100 μm. Images are representative of a minimum of three embryos per genotype. l–o Heat maps of the 40 most significant differentially expressed genes in RNA-Seq analysis of the caudal region of l Axd/Axd or m Grhl2^−/− embryos and their wild-type littermates at E9.5. Boxes represent mean of triplicate samples. Grhl2 over-expression predominantly causes upregulation (yellow shading in l), whereas loss of Grhl2 causes downregulation (blue shading in m). n Among genes upregulated in Axd/Axd and downregulated in Grh2^−/−, 22 were common. o Heat-map of the expression profile of the 22 common genes. Boxes represent individual samples. Source data are provided as a Source Data file