FAM207A subcellular expression and BioID system.
A: Subcellular fractionation of HEK293 Flp-In T-REx cells expressing FLAG-FAM207A. Cy—cytoplasm; Nu—nucleus; Ch—chromatin. B: Western analysis of BirA* constructs used for the FAM207A BioID experiment. Duplicate (induced) lanes are shown. C: Immunolabeling experiment showing co-localization of FAM207A and the IMP3 nucleolar protein. Scale bar = 4μm. D: Dot plot showing prey proteins identified with FAM207A-BirA* that were enriched over endogenous biotinylation (untransfected), unspecific pan-cellular biotinylation (BirA*), and unspecific nuclear biotinylation (NLS-BirA*; BFDR ≤ 5%, SAINT ). Data represent two biological replicates. Proteins in boldface remained statistically enriched when the nuclear localization signal (NLS)-BirA* control was used to further filter the FAM207A BioID data. E: Full western blots for Fig 2A. The red boxes indicate the areas shown in the main figure.