Fig. S3

Localization and long-range signalling activity of Spaw in the presence of FurinA. (A) Extracellular mature Spaw-eGFP can be observed in the LPM of wt embryos (upper row, white arrows). In Zaoh^-/-, no extracellular Spaw-eGFP fluorescence can be detected. (B) Schematic of the transplantation procedure. Donor embryos were injected at 1-cell stage with mRNA of a spaw variant and furina depending on the experiment. At sphere stage, approximately 50 cells were isolated from the donor embryos and transplanted into the animal pole of sphere stage recipient embryos, which were subsequently left to grow for approximately 75 min. (C) After fixation ntla expression was determined by in situ hybridisation (purple staining) and the localisation of the transplanted cells by immunolabelling (brown staining). Expression pattern classes resulting from increasing ntla induction were defined similar to described in (Muller et al., 2012) and used for quantification of the results shown in (D).