Figure S4. Direct interaction of Slug with PIAS family members. A pull-down assay was used to determine the physical interaction between Slug and PIAS family members. Recombinant GST and GSTâSlug proteins were produced from bacteria, and the translated products of HA-tagged PIAS family member genes were obtained using an in vitro transcription/translation system. The production of these proteins was demonstrated by immunoblotting using anti-GST and anti-HA antibodies, respectively. GSTâSlug was used in the pull-down assay for in vitro interaction with HA-tagged PIAS family members. The GST protein alone was used as a negative control. (PDF 24 kb)