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. 2020 Dec 2;162(2):bqaa221. doi: 10.1210/endocr/bqaa221

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© The Author(s) 2020. Published by Oxford University Press on behalf of the Endocrine Society.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 3. — Raloxifene treatment decreases lysine-48 ubiquitinated proteins, activate proteasome assembly, and modulates the expression of DUBs. (A) Western blot analysis for Ub-K48 linked proteins in spinal cord lysate from female c57BL/J6 mice. (B) Quantification of panel A. *P < .05. (C) Western blot analysis for Ub-K48 linked proteins in spinal cord lysate from male c57BL/J6 mice. (D) Quantification of panel C. *P < .05. (E) Venn diagram of genes uniquely upregulated by raloxifene in females or estrogen in males or both. (F) Processes uniquely upregulated by raloxifene in panel E (top 10 hits displayed). (G) Native gel followed by Western blot for the β5 proteasome subunit to distinguish unassembled catalytic core (CP) and assembled with regulatory core (RP) in spinal cord lysates from female c57BL/J6 mice. (H) Quantification of panel G. *P < .05. (I) Native gel followed by Western blot for the β5 proteasome subunit to distinguish unassembled catalytic core (CP) and assembled with regulatory core (RP) in spinal cord lysates from male c57BL/J6 mice. (J) Quantification of panel I. *P < .05. (K) Differential gene expression (log2 fold change) of K48 deubiquitinases (DUBs) compared to vehicle control in female c57BL/J6 mice following raloxifene treatment. Solid bars indicated significant (P < .05) changes vs vehicle control. (L) Differential gene expression (Log2 fold change) of K48 deubiquitinases (DUBs) compared to vehicle control in male c57BL/J6 mice following estrogen treatment. Solid bars indicated significant (P < .05) changes vs vehicle control. (M) Western blot analysis of very high molecular weight (>400 kDa) Ub-K48 linked proteins in spinal cord lysate from female c57BL/J6 mice. (N) Quantification of panel M. *P < .05. (O) Western blot analysis of very high molecular weight (>400 kDa) Ub-K48 linked proteins in spinal cord lysates from male c57BL/J6 mice. (P) Quantification of panel O. *P < .05.