Author: Nila Roy Choudhury; Gregory Heikel; Maryia Trubitsyna; Peter Kubik; Jakub Stanislaw Nowak; Shaun Webb; Sander Granneman; Christos Spanos; Juri Rappsilber; Alfredo Castello; Gracjan Michlewski
Title: RNA-binding activity of TRIM25 is mediated by its PRY/SPRY domain and is required for ubiquitination Document date: 2017_10_9
ID: ifla4aix_101
Snippet: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/200410 doi: bioRxiv preprint IPed T7-TRIM25, T7-TRIM25ΔRING or T7-TRIM25K117R from HeLa cell extracts in the presence of recombinant purified 6xHis-Ube2D3, UBE1 and ubiquitin. Lanes 1, 2 and 3 show IPed T7-tagged proteins. Lanes 4, 6 and 8 represent 60 minutes' reaction with T7-tagged IPed proteins, 6xHis-Ube2D3, UBE1 and ubiquiti.....
Document: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/200410 doi: bioRxiv preprint IPed T7-TRIM25, T7-TRIM25ΔRING or T7-TRIM25K117R from HeLa cell extracts in the presence of recombinant purified 6xHis-Ube2D3, UBE1 and ubiquitin. Lanes 1, 2 and 3 show IPed T7-tagged proteins. Lanes 4, 6 and 8 represent 60 minutes' reaction with T7-tagged IPed proteins, 6xHis-Ube2D3, UBE1 and ubiquitin. Lanes 5, 7 and 9 represent 60 minutes' reaction with T7-tagged IPed proteins, UBE1 and ubiquitin. The proteins were analyzed by western blotting with anti-TRIM25 antibody. HeLa cell extracts overexpressing T7-TRIM25 or T7-TRIM25ΔRBD. Lanes 1, 4 and 7 represent 4% (40 μ g) of the loading controls (input). Lanes 2, 5 and 8 represent beads only All rights reserved. No reuse allowed without permission.
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