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. 2022 Mar 22;23(7):3421. doi: 10.3390/ijms23073421

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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NPM1c expressing cells exhibit low basal levels of SUMOylation, and EAPB0503 restores NPM1c post-translational modifications triggering its proteasomal degradation. (A) Western blot analysis of NPM1c and Actin in OCI-AML3 and OCI-AML2 cells treated with EAPB0503 for 6, 24 and 48 h. Histogram represents the average densitometry of NPM1c/Actin in OCI-AML3 cells in three independent experiments. (B) Endogenous interactions detected by Duolink between NPM1 (wt + c) and SUMO 2/3 (Red) in untreated OCI-AML3 and OCI-AML2 cells (upper panel) or EAPB0503 treated cells for 6 h (lower panel). Nuclei were stained with 4-,6-diamidino-2-phenylindole (DAPI) (blue). (C) OCI-AML3 and OCI-AML2 cells were treated with EAPB0503 and PS-341 for 6 h. NPM1 (wt + c) immunoprecipitates [IP-NPM1 (wt + c)] were blotted for SUMO 2/3. Input was blotted for NPM1c, NPM1 (wt + c) and H3. (D) OCI-AML3 and OCI-AML2 cells were treated with EAPB0503 and PS-341 for 24 h. NPM1 (wt + c) immunoprecipitates [IP-NPM1 (wt + c)] were blotted for Ubiquitin. Input was blotted for NPM1c, NPM1 (wt + c) and H3. *** p < 0.001.