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. 2021 Jun 24;9:693079. doi: 10.3389/fcell.2021.693079

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Copyright © 2021 Zhang, Chen, Sun, Zhang, Zhang, Li, Huang, Yang and Ye.

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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Peroxisome proliferator-activated receptor γ2 (PPARγ2) can be SUMO2/3 modification during adipogenic induction and deSUMOylated by SENP3. (A) The endogenous SUMO2/3 modification of PPARγ2 in BM-MSCs during adipogenesis. Whole-cell lysates were prepared from BM-MSCs undergone adipogenic differentiation at day 3 and were incubated with antibody specific to PPARγ2. The co-eluted proteins were detected by using PPARγ2 and SUMO2/3 antibody. (B) PPARγ2 can be modified by SUMO3 when co-expressed in HEK293T cells in dose-dependent manner; Flag-PPARγ2, ubc9 and RH-SUMO3 were co-expressed in HEK293T cells and co-IP assays were performed with Flag and RGS antibody; (C) the SUMOylation of PPARγ2 can be removed by SENP3; Flag-PPARγ2, ubc9, Flag-SENP3, and RH-SUMO3 were co-expressed in HEK293T cells and co-IP assays were performed with Flag and RGS antibody; (D) SENP3 mediated de-SUMOylation depended on its active site cysteine 532 (C532); Flag-PPARγ2, ubc9, Flag-SENP3 (WT and C532A mutant) and RH-SUMO3 were co-expressed in HEK293T cells and co-IP assays were performed with Flag and RGS antibody.