Fig. 5. The V62E mutation in BRAT1-mutated patient cells impairs BRAT1 interaction with INTS11 and INTS9.

a Western blot of BRAT1, INTS11 and INTS9 levels in BRAT1 immunoprecipitates from control, parent, BRAT1 patient-derived fibroblasts and lymphoblastoid cell lines (LCLs), as indicated. Note that 2-fold more lysate from patient cells was employed for loading (input) and immunoprecipitation to ensure equivalent levels of INTS11. The black asterisk marks a nonspecific band. b BRAT1^−/− (clone #8) cells transiently transfected with FLAG-tagged BRAT1^WT and BRAT^V62E constructs were fixed 24 h post-transfection and stained with antibody against FLAG-tag (green) and DAPI (blue). c Levels of BRAT1, INTS11, INTS9 and FLAG-tagged BRAT1 in BRAT1 (left) and FLAG (right) immunoprecipitates from BRAT1^−/− (clone #8) cells transiently transfected with FLAG-BRAT1^WT or FLAG-BRAT^V62E, measured by Western blotting. a–c The experiment was performed three times with similar results. Uncropped and unprocessed scans are provided in the Source Data file.