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ATRX BioID system and proximal associations.

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journal contribution
posted on 15.11.2021, 18:42 by William A. Scott, Erum Z. Dhanji, Boris J. A. Dyakov, Ema S. Dreseris, Jonathon S. Asa, Laura J. Grange, Mila Mirceta, Christopher E. Pearson, Grant S. Stewart, Anne-Claude Gingras, Eric I. Campos

A: Western blot exemplifying stable integration and expression of FLAG-tagged BirA* protein fusions in inducible HEK293 Flp-In T-REx cells (top), and similar expression levels of N- and C-terminally-tagged ATRX (bottom). B: Western blot (top) and Coomassie-staining (bottom) of biotinylated proteins captured on streptavidin beads. C: Example of immunofluorescent labeling demonstrating nuclear targeting of the ATRX-BirA* fusion constructs. D: Gene ontology analysis of ATRX-associating proteins. E: Western blot showing the subcellular distribution of ATRX-associating proteins in HeLa S3 cells. F: Examples of the proximity ligation assay (PLA) showing associations between endogenous proteins in MO3.13 cells. G-H: PLAs showing associations between endogenous ATRX and exogenous FLAG-tagged CCDC71 (G) or FAM207A (H) in HEK293 Flp-In T-REx cells. I: PLAs showing an association between endogenous ATRX and myc-tagged SLF2. Data plots account for ∼100 nuclei in three independent experiments, with the total number of nuclei assessed in brackets. The p-values were obtained using a 2-sided Student’s t-test with unequal variance. J: In vitro interaction between recombinant FAM207A and ATRX.