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. 2022 Oct 3;13(10):844. doi: 10.1038/s41419-022-05292-9

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© The Author(s) 2022

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. 2 — A Knockdown of NAE1 or UBA3 had a minor effect on PD-L1 levels. H358 and BxPC3 cells were transfected with siRNAs targeting NAE1, UBA3 or control siCont for 48 h, followed by IB with the indicated Abs. B NAE1 or UBA3 knockdown had a minor effect on MLN4924-induced PD-L1 induction. H358 cells were transfected with indicated siRNAs for 24 h, then cells were treated with the indicated concentrations of MLN4924 for 24 h, followed by IB with the indicated Abs. C Proteasome inhibitor MG132 had minor effect on PD-L1 levels. H358 cells were treated with MLN4924 (0.5 μM) for 24 h or MG132 (10 μM) for 8 h or MG132(1 μM) for 24 h, followed by IB with the indicated Abs. D MLN4924 induced the phosphorylation of ERK1/2, AKT, JNK and c-FOS in dose- and time-dependent manners. H358 cells were treated with various concentrations of MLN4924 for the indicated time periods, followed by IB with the indicated Abs. E MLN4924-induced elevation of both PD-L1 protein (Top) and mRNA (Bottom) can be largely abrogated by MEK inhibitor (trametinib) or JNK inhibitor (SP600125). H358 cells were treated with either MLN4924 or indicated inhibitors or both, followed by IB with the indicated Abs. F MLN4924 induction of both PD-L1 protein (Left) and mRNA (Right) can be largely abrogated by knockdown of ERK1/2, or c-JUN. H358 cells were transfected with siRNAs targeting ERK1/2, c-JUN or control siRNA, followed by treatment with MLN4924 (0.5 μM) for 24 h, then for IB with the indicated Abs. G MEK inhibitor-induced abrogation of PD-L1 induction occurred at the transcriptional level. H358 cells were treated with MLN4924 alone or combined with indicated inhibitors, followed by IB with the indicated Abs. MEKi MEK inhibitor (trametinib), JNKi JNK inhibitor (SP600125), AKTi AKT inhibitor (MK2206), MG132 proteasome inhibitor, CQ (chloroquine) lysosome inhibitor, se short exposure, le long exposure.