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. 2015 Nov 17;6(6):e01777-15. doi: 10.1128/mBio.01777-15

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Copyright © 2015 Bowman et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

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FIG 3 — HOIP is ubiquitinated upon TLR4 stimulation. (A) A20.2J mouse B cells were stimulated with LPS and lysed under denaturing conditions. Endogenous HOIP was immunoprecipitated, separated by SDS-PAGE, and analyzed by immunoblotting. High-molecular-weight HOIP bands (>120,000) appeared after LPS stimulation. (B) Complemented mouse A20.2J HOIP^−/− cells used for Fig. 2C were stimulated with LPS for the indicated times and lysed under denaturing conditions. Anti-IgG (control)- or anti-Flag (HOIP)-immunoprecipitated samples were prepared as described for panel A. A long exposure of the Flag immunoblot revealed the presence of high-molecular-weight HOIP bands. Data are representative of three independent experiments.