Figure 1—figure supplement 2.

Characterization of the reck^ulb3 allele.

(A) Schematic representation of the domain architecture of Reck, with from N- to C-terminus, five cysteine knot (CK) motifs, a Frizzled-like cysteine-rich domain (CRD), two epidermal growth factor-like domains (EGF-like) and three Kazal-like motifs upstream of a membrane glycosylphosphatidylinositol (GPI) anchor. The CK motifs are implicated in Wnt signaling by binding Gpr124 and Wnt7a/b, while the Kazal motifs control matrix metalloproteinase (MMP) activity. (B) Sequence alignment of wild-type (WT) and mutant Reck sequences, showing the disulfide bonding pattern within the CK motif consensus (6 Cys repeat: C-C-X7-9-C-X3-C-X12-22-C-X9-12-C). The reck^ulb3 mutant allele was generated by CRISPR-Cas9 mutagenesis at the target site highlighted in blue (see Materials and methods for details). PAM: protospacer adjacent motif. An in-frame mutation allele was selected to interfere selectively with the domains of Reck implicated in Wnt-dependent brain angiogenesis.