Figure 5.

Casp8 physically interacts with AR. (A) Casp8 interacts with multiple regions in AR. GST, GST-AR (1–36), GST-AR (334–566), or GST-AR (662–919) was immobilized on glutathione-Sepharose beads and incubated with ³⁵S-labeled Casp8. Proteins retained on the beads were separated by SDS–PAGE and the gel was analyzed by autoradiography. Five percent of input (IP) for the interaction assay was loaded on the same gel. (B–D) Identification of motifs that mediate Casp8–AR interactions. GST, GST-AR (1–36), GST-AR (1–36/F26A/F27A), GST-AR (334–566), GST-AR (334–566/L436A/F437A), or GST-AR (662–919) was immobilized on glutathione-Sepharose beads and incubated with ³⁵S-labeled Casp8 (1–43), Casp8 (1–49), Casp8 (1–49/F45A), Casp8 (200–290), Casp8 (200–300), Casp8 (200–300/F295A/I296A), Casp8 (1–170), Casp8 (1–180), or Casp8 (1–180/I174A/I175A) as indicated. After being washed, beads were submitted to SDS–PAGE and the gel was analyzed by autoradiography. The input (IP) (5%) for the interaction assay was loaded onto the same gel. (E) Diagram of motifs in AR and Casp8 that are involved in Casp8–AR interactions. Black boxes represent motifs that mediate Casp8–AR interactions.