RDH10 Control of RA Synthesis Figure 4. Phenotype and characterization of transcript of Rdh10delta allele. (A,C) Wild type and (B, D) homozygous Rdh10delta/delta embryos were collected at E9.5 (A–B) and E10.5 (C–D. (E9.5 mutant embryos n = 34; rightward turn L complete n = 33. E9.5 with leftward turn n = 1. E10.5 mutant embryos n = 5). Formalin-fixed embryos were stained with DAPI and imaged by confocal microscopy. For each embryo, a Z-stack of confocal slices was collapsed to form a ‘‘pseudo-SEM’’ image. (E) Schematic diagram of Rdh10 spliced mRNA exon structure, along with coding sequence for Rdh10 wild-type (full length) and Rdh10delta mutant (truncated) as determined by direct sequencing of reverse transcribed Rdh10 mRNA. Yellow boxes indicate spliced exons of Rdh10 mRNA. Dark blue arrow represent coding sequence of spliced wild type Rdh10 mRNA, blue boxes beneath arrow symbolize truncated coding sequence resulting from splicing of exon 1 directly to exon 3. Nucleotide and protein sequence indicated beneath schematic diagram shows 39 end of exon 1 spliced directly to 59 end of exon 3, generating a premature stop codon. Pink box indicates exon 1 derived sequence, green box represents exon 3 derived sequence. (F) Histogram of elative level of Rdh10 mRNA as assessed by reverse transcription qPCR. doi:10.1371/journal.pone.0030698.g004 PLoS ONE | www.plosone.org 6 February 2012 | Volume 7 | Issue 2 | e30698