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. 2021 Jun 7;9:684885. doi: 10.3389/fcell.2021.684885

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Copyright © 2021 Gong, Wang, Wang, Hu, Li, Gao and Lin.

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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ASB6 interacts with p62 and it is part of CRL5 E3 ligase complex. (A,B) HEK293T cells transfected with the indicated plasmids were treated with 10 μM MG132 for 10 h before harvest for Co-IP and IB analysis. (C) Schematic illustration of human ASB6 protein domain composition. (D,E) HEK293T cells were co-transfected with ASB6 wild type and mutants (Δ-SOCS box, Δ-Cul5-box) and HA-CUL5 (D) or HA-p62 (E) constructs. Cells were treated with 10 μM MG132 for 10 h before harvest for Co-IP and IB analysis. (F) Huh1 cells were treated with 10 μM MG132 for 10 h before harvest to make whole-cell lysates. ASB6 antibody was used to perform endogenous Co-IP experiments with Rabbit IgG as negative control.