Figure 7.
Co-IP of in vitro-translated FANCA and FANCG. Proteins were cotranslated in vitro and labeled with [35S]methionine. (A) Full-length FANCA (1–1455) and carboxyl terminal-truncated FANCA mutants (1–1200, 1–900, 1–600, and 1–300) were immunoprecipitated with a rabbit polyclonal antibody directed against the amino terminus of FANCA (17). After SDS/PAGE, coprecipitating HA-FANCG was visualized by exposing the dried gel to x-ray film. (B) HA-FANCG was immunoprecipitated with anti-HA to identify bound FANCA or carboxyl terminus-deleted forms of FANCA as described in A. (C) Complex formation was examined between HA-FANCG and FANCA 342-1455 (Left) or FANCG and FANCA 342-1455-HA (Right) after IP with anti-HA. Total lysate (TL) shows the presence of FANCG (arrows) and amino terminal-deleted FANCA (arrowhead). Immunoprecipitated products are shown in the lanes labeled IP.