Fig. 6.

Identification of the binding regions that are critical for Nbs1-Smad7 interaction. a GST-tagged Smad7 was transfected along with a 3HA-tagged Nbs1 fragment. Whole cell lysates were pulled-down with the HA-specific antibody and the interacting Smad7 was detected using an anti-GST antibody. b To identify the specific binding region of Nbs1 that is important for its interaction with Smad7, Nbs1 fragments were immunoprecipitated with Glutathione Sepharose 4b beads, and the interacting Smad7 was detected using the Flag-specific antibody. c, d To identify the region of Smad7 that is important for its interaction with Nbs1, all Smad7 fragments described in Online Resource 8 were immunoprecipitated using Glutathione Sepharose 4b beads, and the interacting Nbs1 was detected using the HA-specific antibody