Fig. 5

The cLEN repressive element, but not a neighboring sequence, is present in other Danio species. (A) Alignment of orthologous sequences of cLEN11-1 to cLEN11-5 in Danio species, including Danio rerio, Danio aesculapii, Danio kyathit and Danio albolineatus. cLEN-22 contains 6 or 9 bp insertions in non-zebrafish sequences. Asterisks indicate conserved base pairs. The conserved GTCA sequence overlapping cLEN-11-1 and cLEN-11-2 is marked in red. Dendrogram indicates phylogenetic relationship. (B) Images of cardiac sections of cLEN^ins6bp:EGFP transgenic fish harboring a 6 bp insertion (TGTGTA). (C) Images of cardiac sections of cLENm^gtca:EGFP transgenic fish harboring the mutation of GTCA to CATT. The arrows indicate endocardial EGFP expression. Numbers of animals are shown in Table S3. (D) RT-qPCR analysis of lepb in uninjured and 3 dpa hearts of D. aesculapii. (E) RT-PCR of samples from the uninjured and 3 dpa hearts of D. kyathit. actb2 was used as the loading control. Uninj., uninjured hearts. (F) RT-qPCR analysis of lepb in the uninjured hearts of zebrafish, D. aesculapii, D. kyathit and D. albolineatus. The data are presented as mean±s.d., n=3; lepb transcript levels were normalized to actb2 levels in D,F. *P<0.05; N.S., not significant. Student's unpaired, two-tailed t-test. Scale bars: 100 µm in B,C.