Figure 5.

SLIT2 modulates N-cadherin (N-Cadh)/β-catenin (β-cat) signaling to influence Schwann cell migration ability. (a) N-cadherin/ β-catenin binding was analyzed by co-immunoprecipitation (IP) in SNF cells after incubation with F+M (control) or FcoM media. (n=3). (b) N-cadherin/β-catenin binding was analyzed by co-immunoprecipitation in SNF cells after incubation with F+M (control) or FcoM media from fibroblasts transfected with Ctr (Si-Ctr) or SLIT2 (Si-SLIT2) siRNA. (n=3). (c) Nuclear extracts from SNF cells incubated with SNF, F+M or FcoM media (with fibroblasts transfected with Si-Ctr or Si-SLIT2 siRNA) were analyzed for β-catenin expression. Lamin A/C was used as a loading control and β-tubulin was used asa quality control of nuclear extracts. β-Catenin expression was normalized to respective lamin A/C expression (n=3). (d) mRNA level of three β-catenin targets (C-MYC, LCF4 and MMP9) analyzed by quantiative real-time PCR (QRT-PCR) in SNF cells after incubation with SNF (used as normalizer), F+M or FcoM media (n=3; *P<0.05; **P<0.01; ***P<0.001). Md, conditioned media from: SNF, sNF 96.2; F, fibroblasts; M, macrophages; F+M, mixed Md from separated cultures of fibroblasts and macrophages; FcoM, cocultures of fibroblasts and macrophages