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. 2019 Nov 14;9:16819. doi: 10.1038/s41598-019-53418-5

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© The Author(s) 2019

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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Characteristics of rat primary-cultured peripheral blood mononuclear cells (PBMCs) preconditioned by oxygen-glucose deprivation (OGD) (OGD-PBMCs). The levels of the secretory vascular endothelial growth factor (VEGF) (N = 4) (A), anti-inflammatory cytokine transforming growth factor-β (TGF-β) and pro-inflammatory cytokine tumour necrosis factor-α (TNF-α) (N = 7 each) (B) from the conditioned media of rat primary-cultured PBMCs and isolated monocytes subjected to either normoxia (norm) or OGD. Western blotting for TNF-α showed a band around 42 kDa, a weak 25 kDa band and an 18 kDa band (arrow). (C) The ratio of TGF-β per TNF-α, which reflects the PBMCs polarisation, under both the normoxic and OGD conditions (N = 7 each). The increase in the ratio of TGF-β per TNF-α reveals the polarisation to the protective anti-inflammatory status under the OGD condition. (D) Representative figures indicate the expression of the pro-inflammatory marker (i.e., inducible nitric oxide synthase, iNOS) and the protective anti-inflammatory marker (cluster of differentiation 206, CD206) in the cell lysate from PBMCs under both the normoxic and the OGD conditions (N = 6 each). (E) The ratio of CD206 per iNOS, which reflects the PBMC polarisation under both the normoxic and OGD conditions (N = 6 each). The increase in the ratio of CD206 per iNOS reveals the polarisation of the protective anti-inflammatory status under the OGD condition. (F) Representative figures indicate the expression of the peroxisome proliferator-activated receptor γ (PPARγ) in the cell lysate from PBMCs under both the normoxic and the OGD conditions (N = 6 each). Transferrin and β−actin confirmed an equal loading of proteins. Data represent the relative optical densities of each sample compared to the loading control band by unpaired t-test. ^*P < 0.05. ^**P < 0.01.