Fig. S6

DFC specification and cell number were not altered in lpar3 and atx morphants. (A-I) Wild-type (Wt) embryos, lpar3 and atx morphants were subjected to WISH against the DFC-specific marker ntl (A-F) at 60% epiboly and the six-somite stage (6 SS) and lrdr1 + lefty1 (G-I) at the ten-somite stage (10 SS; as indicated). Formation of DFCs and sustained expression of DFC-specific genes were observed in Wt embryos as well as in lpar3 and atx morphants at the stages indicated (open arrowheads). Notochord expression of both ntl and lefty1 were also observed (filled arrowheads). (J-O) Embryos were stained with vital dye SYTO-11 (10 mM) and examined at 6.5 hpf for DFCs endocytosis activity, cell clustering and cell number quantification. DFCs in lpar3 (K,N) and atx (L,O) morphants formed as early as those in Wt embryos (J,M), with similar numbers, but are more dispersed as linear pattern and sometimes even separate into two groups.