Figure 3.

p107^FHIP interacts with FTS and Hook proteins to form the FHF complex. (A) GST-p107^FHIP interacts with FTS and Hook1. Plasmids expressing the indicated FLAG-tagged wild-type or mutant FTS proteins were transfected into HEK293T cells together with plasmids expressing either GST-p107^FHIP or GST and after 48 h, cell extracts were generated and incubated with GSH-Sepharose. Bound proteins and control lysates were subjected to immunoblotting with the indicated antibodies. (B–D) HEK293T cells stably expressing FLAG-HA-Hook3 were generated, and extracts were examined by immunoblotting using antibodies against Hook3 (B). Anti-HA or control immune complexes from HEK293T/FLAG-HA-Hook3 cells were immunoblotted with the indicated antibodies (C) or eluted with HA peptide before trypsinization and LC/MS/MS analysis (D). The number of independent peptides as well as the total number of scans for these peptides is indicated, as well as the coverage of each protein.