Fig. 5

Expression validation of concordant Shh-regulated genes. a-b Vehicle-normalized expression relative to Gapdh is shown for the top 10 (a) up- and (b) downregulated genes with concordant expression changes. Mean fold change (SHH/Veh) +/− SEM is shown. Known orofacial cleft-associated genes in humans are highlighted in magenta [15]. N = 4 samples per treatment group. c Pseudo-colored scanning electron micrographs are shown for an intact and a sagittally cut (white dashed line) GD11 mouse embryo. The cranial neural crest-derived mesenchyme (orange) and surface ectoderm (teal) of the MNPs of GD10 or GD11 embryos were separated and isolated for RT-PCR analysis. d-e Tissue-specific expression enrichment was determined for the top 10 up- and downregulated concordant genes. Values represent the mean expression per Gapdh for the mesenchymal (orange) and ectodermal (teal) compartments of the MNPs for each gene divided by the total expression in both tissues multiplied by 100%. Sox10 (orange) is a specific marker of the cNCC-derived mesenchyme, and Cdh1 (E-cadherin, teal) is a specific marker of the surface ectoderm. Surface ectoderm and mesenchyme tissues were isolated from N = 3 pooled litters per gestational stage. MNP = medial nasal process, LNP = lateral nasal process, MxP = maxillary process, M = mesenchyme, E = ectoderm, OFC = orofacial cleft