Fig. 2

s1pr1 overexpression affects heart and fin development in a dose-dependent way. Analysis of 72 hpf Tg(Myl7:EGFP) embryos injected with CDS or CDS+UTR mRNA of s1pr1at the reported doses (pg). The percentage of embryos with the indicated heart (A) or pectoral fin (D) defects was averaged across multiple independent experiments carried out in double blind. The total number of analyzed embryos were as follows: RFP mRNA injected n = 185 (50 pg), n = 138 (100 pg), n = 89 (200 pg); s1pr1CDS mRNA injected n = 195 (50 pg), n = 199 (100 pg), n = 106 (200 pg); s1pr1 CDS+UTR mRNA injected n = 214 (50 pg), n = 198 (100 pg), n = 96 (200 pg). For all tested doses, differences between control and s1pr1 injected embryos were significant (Fisher's test P < 0.0001) with the only exception of the 50 pg mRNA CDS+UTR which was not significant. (B,C,E) Images representative of the different phenotypes. In (B), the red dashed arrows emphasize the valve orientation as indicator of correct looping. In (C) an example of cardia bifida phenotype is shown. In (E) the yellow arrow indicates the absence of fins, the green arrows indicate WT fins and the pale green arrow points to an example of defective fin. V, ventricle; a, atrium. Red scale bar = 100 μm.