Fig. 1

Sequence and expression analysis of ccm2l. (A) Alignment of Ccm2l and Ccm2 protein sequences. (B) Schematic comparing the domain structures of Ccm2l and Ccm2. The N-terminal region of Ccm2l contains a putative PTB domain with high identity to the PTB domain of Ccm2. The two proteins also have regions of high identity at their C-termini. (C) RT-PCR showing expression of ccm2, ccm2l, ccm1 and the positive control gapdh at five embryonic stages. All four genes are expressed at all five time points tested between 0 and 48 hpf. Water (w) is included as a negative control. (D) RT-PCR comparing expression of ccm2l and ccm1 in whole embryos (e) and purified embryonic hearts (h) at 48 hpf. Both genes are expressed in the heart, with the ratio of heart expression to whole embryo expression much higher for ccm2l. RT-PCR for the heart-specific marker cardiac myosin light chain (cmlc2) and for a transcript absent from the heart, insulin (ins), demonstrate the quality of the heart tissue purification. (E) In situ hybridization analysis of ccm2l expression. In 30 hpf embryos, ccm2l expression is detected in the posterior end of the embryo in the presumptive notochord (arrowheads) and the tissue ventral to it (arrows). Embryos treated with sense probe as a negative control lack staining in these domains. PH, Pleckstrin-homology superfamily; PTB, phosphotyrosine binding domain; w, water.