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. 2018 Jan 9;8:217. doi: 10.1038/s41598-017-18641-y

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© The Author(s) 2017

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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Effects of SEP and αPD-L1 combination on MEK/ERK signaling pathway in T lymphocytes in vitro. The T cells from naïve mice were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum. After the cells were pretreated with/without MEK inhibitor PD98059 (40 μM) for 30 min, they were cultured with the aPD-L1 (10 nM) and/or SEP (50 μg/ml) with ConA (2 μg/ml) for additional 48 h. The T cells were collected and lysed in lysis buffer supplemented with a cocktail of protease and phosphatase inhibitors. Western blot analysis was conducted for the detection of P-ERK1/2 and ERK1/2 expression. αPD-L1 or SEP in combination with ConA significantly increased the ratios of P-ERK1/2 and ERK1/2 compared with ConA group (P < 0.05, respectively, n = 3). Furthermore, combination of SEP and αPD-L1 had more potent than SEP (P < 0.05, n = 3) or αPD-L1 (P < 0.05, n = 3). However, after the pretreatment with PD98059 for 30 min, the the increased ratios induced by SEP or/and αPD-L1 was partially decreased (P < 0.05, respectively, n = 3). The results were presented as mean ± SEM. ^a P < 0.05 compared with control group. ^b P < 0.05, compared with SEP and αPD-L1 combination group (group 4). ^c P < 0.05, compared with SEP and αPD-L1 combination group after PD98059 pretreatment for 30 min (group 7). ^* P < 0.05, group 2 compared with group 5; group 3 compared with group 6; group 4 compared with group 7.