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. 2019 Jul 19;10:1495. doi: 10.3389/fimmu.2019.01495

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Copyright © 2019 Chapman, Corridoni, Shiraishi, Pandey, Aulicino, Wigfield, do Carmo Costa, Thézénas, Paulson, Fischer, Kessler and Simmons.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Ubiquitome screen reveals novel DUB targets of ataxin-3. (A) Flow chart showing experimental protocol. Control and ataxin-3 shRNA THP-1 cells were left unstimulated or stimulated for 60 min with MDP + PAM₃CSK₄ prior to ubiquitin enrichment using TUBEs1 beads, with the lysates subsequently subjected to LC-MS/MS analysis. (B) Immunoblot using antibody against K63 linked polyubiquitinated proteins of whole cell lysates (input) and TUBEs1 enriched fractions or beads only control samples. Volcano plots comparing ubiquitinated protein expression in (C) unstimulated control and ataxin-3 shRNA THP-1 cells and (D) control and ataxin-3 shRNA THP-1 cells following MDP + PAM₃CSK₄ stimulation. The dotted lines on the y-axes represents a p-value of 0.05 (t-test), and on the x-axes represents fold change >2 or <-2.