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PxVxL motif in CCDC71L, CCDC71 subcellular fractionation and immunoprecipitation, and BioID system.

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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: PxVxL motif in human CCDC71L. B: Subcellular fractionation of HEK293 Flp-In T-REx cells expressing FLAG-CCDC71. Cy—cytoplasm; Nu—nucleus; Ch—chromatin. C: CCDC71 immunoprecipitation and verification of associations reported in BioGrid [52]. D: Western analysis of CCDC71 and control BirA* constructs. E: Dot plot showing prey proteins identified with CCDC71-BirA* that were enriched over endogenous biotinylation (untransfected) and unspecific pan-cellular biotinylation (BirA*; BFDR ≤ 5%, SAINT [103]). 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 CCDC71 BioID data. F: Full western blots for Fig 3D. The red boxes indicate the areas shown in the main figure.

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