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. 2015 Aug 10;290(39):23850–23862. doi: 10.1074/jbc.M115.664375

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© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 8. — NUB1L overexpression accelerates the turnover of ubiquitinated misfolded proteins. A, the effects of NUB1L on proteasome abundance. Western blots of the indicated proteasome subunits in total cell extracts are shown. B, the effects of NUB1L on proteasome peptidase activities. n = 6 biological repeats. N.S., not significant. C and D, NRVMs were infected with Ad-NUB1L or Ad-Gal for 72 h. Ad-HA-CryAB^R120G was infected 24 h prior to Ad-NUB1L infection, and BZM was added 24 h before cell harvesting. C, NUB1L overexpression reduced Lys-48-linked ubiquitinated CryAB^R120G, and proteasome inhibition attenuated the reduction. CryAB^R120G was immunoprecipitated (IP) with anti-HA antibodies from cell lysate and blotted (IB) with the indicated antibodies. Data are representative of three independent repeats. D, polyubiquitinated proteins were precipitated from cell lysates using TUBEs under denaturing conditions and blotted with the indicated antibodies. Data are representative of three independent repeats.