Figure 5.
Figure 5. Interaction of FANCA mutants with the FANCB/FANCL complex and with FANCG. Stable cell lines were generated expressing FANCA protein containing patient-derived missense mutations and immunoprecipitation with an α-FLAG affinity gel (A-C), or an antiserum against the N-terminus of FANCA (D) was performed. Western blots were probed for FANCA (Rb89), FANCG (Rb43), and FANCL.12 The FANCG protein is indicated with an asterisk. (E-G) Mammalian 3-hybrid assay in 293 cells expressing FLAG-FANCB. Cells were cotransfected with the indicated protein pairs and assayed for luciferase activation. Fold induction relative to reporter gene activation when full-length FANCL is expressed alone. (E-G) Error bars represent SD of 3 experiments.

Interaction of FANCA mutants with the FANCB/FANCL complex and with FANCG. Stable cell lines were generated expressing FANCA protein containing patient-derived missense mutations and immunoprecipitation with an α-FLAG affinity gel (A-C), or an antiserum against the N-terminus of FANCA (D) was performed. Western blots were probed for FANCA (Rb89), FANCG (Rb43), and FANCL.12  The FANCG protein is indicated with an asterisk. (E-G) Mammalian 3-hybrid assay in 293 cells expressing FLAG-FANCB. Cells were cotransfected with the indicated protein pairs and assayed for luciferase activation. Fold induction relative to reporter gene activation when full-length FANCL is expressed alone. (E-G) Error bars represent SD of 3 experiments.

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