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Fig. 1. Targeted truncation of ROR2 at W749. (A) Structure of the ROR2 receptor showing the immunoglobulin-like (Ig), frizzled-like (Frz), kringle (Kr), transmembrane (TM), tyrosine-kinase (TK), serine/threonine-rich (ST1, ST2) and proline-rich (PR) domains, and the site of the BDB mutation (black line). SP, signal peptide. (B) Targeting strategy in the region spanning the last exon of Ror2, encoding the TK and ST1/PR/ST2 domains. The targeting vector carries a replacement of the region encoding W749-D930 with a FLAG-encoding sequence fused in frame, followed by a selectable marker (NEO, neomycin resistance gene). Target sites for primers are indicated by white (cDNA genotyping) and black (genomic genotyping) arrowheads. (C) PCR analysis of yolk sac DNA from wild-type and mutant embryos showing the products from the wild type (WT, 419 bp) and targeted (T, 174 bp) alleles. (D) RT-PCR analysis confirming the loss of wild type Ror2 mRNA (455 bp product, WT) in Ror2^{W749FLAG/W749FLAG} E15.5 embryos (KI), and the presence of mRNA for the truncated ROR2 (321 bp product, T). (E) Immunoblot analysis of ROR2 receptor from wild-type (WT) and Ror2^{W749FLAG/W749FLAG} (KI) tissues. Antibodies used for immunoprecipitation (IP) and western (W), and sizes (kDa) and migration of molecular standards, are indicated. (F,G) Confocal microscopy of full-length and truncated ROR2 receptors overexpressed in Cos1 cells.

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