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. 2021 Aug 4;10(8):1245. doi: 10.3390/antiox10081245

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© 2021 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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(A) An illustration of the TAT-HA-Prdx6 expression vector system. Constructs i, ii, and iii represent TAT-HA-Prdx6^WT, Sumoylation-deficient TAT-HA-Prdx6^K122/142R, and TAT-HA-Prdx6^{C47IL/S32A/H26A/D140A} (inactive-mutant), respectively. These plasmids were utilized for recombinant protein preparation. (B,C) Expression of TAT-HA-Prdx6^WT, and its mutant TAT-HA-Prdx6^K122/142R and TAT-HA-Prdx6^{Inactive-mutant} fusion recombinant protein in Escherichia coli. (B) TAT-HA-Prdx6^WT or its mutant at K122/142R and inactive-mutant (C47IL/S32A/H26A/D140A) fusion bacterial protein expression was analyzed using Coomassie blue staining of SDS-PAGE gel (upper panel) and immunoblot analysis with anti-Prdx6 antibody (lower panel). (C) Purified TAT-HA-Prdx6^WT and its mutants TAT-HA-Prdx6^K122/142R and TAT-HA-Prdx6^{Inactive-mutant} protein were analyzed using Coomassie blue staining (upper panel) and immunoblot analysis with Prdx6 antibody (lower panel).