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. 2020 Mar 9;11(3):178. doi: 10.1038/s41419-020-2369-4

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a HepG2 cell lysates (1 mg) were incubated with pure TNFAIP8 protein (2 µg) for 3 h. TNFAIP8 was pulled-down with TNFAIP8 antibody or control IgG. Immunocomplexes were resolved by SDS-PAGE and WB with indicated antibodies. b, c Interaction of TNFAIP8-Myc protein with ATG7-ATG3-LC3B complex was analyzed by forward and reverse IP as indicated. d Effect of TNFα on cellular ROS production was visualized by staining the cells with CellROX Green reagent (Invitrogen). e Effect of TNFα on TNFAIP8 and IL-6 mRNA expression was analyzed by RT/qPCR (n = 3). f, g Effect of TNFα on ATG3, ATG7. TNFAIP8 and LC3B protein expression were analyzed by WB and interaction of TNFAIP8 with ATG3-ATG7 was analyzed by TNFAIP8 IP followed by WB. *P < 0.05, **P < 0.01 relative to untreated SK-Hep1 cells. IP: immunoprecipitation, WB: western blotting.