Figure 2. Specific in vitro interactions between EN and GsbN proteins.

A and B: GST-pull down assays, A of S³⁵-radiolabelled full-length GsbN protein with GST (negative control) or GST-EN fusion protein, and B of S³⁵-radiolabelled full-length EN protein with GST (negative control) or GST-GsbN fusion protein. In each case, control of migration corresponds to 1/10 of the input. C and D: Co-immunoprecipitation between EN and GsbN. C Immunoprecipitation of S³⁵-radiolabelled GsbN was first tested (GsbN* IP), showing that guinea pig anti-GsbN was able to retain GsbN protein. Co-immunoprecipitation was performed using S³⁵-radiolabelled EN protein in the presence of protein extracts from HS-GsbN embryos. D Immunoprecipitation of S³⁵-radiolabelled EN was first tested (EN* IP), showing that 4F11 anti-EN was able to retain EN protein. Co-immunoprecipitation was performed using S³⁵-radiolabelled GsbN protein in the presence of protein extracts from HS-EN embryos.