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All samples were analysed on a FACSCalibur flow cytometer using CellQuest software (BD Biosciences). Dendritic cells were isolated from iDLNs of C57Bl/6 http://www.selleckchem.com/products/Everolimus(RAD001).html mice 4?days after one EFD BCG injection. Following cell sorting, pDCs (CD11clow B220high) and conventional DCs (cDCs; CD11chigh B220neg) were cultured alone or co-cultured with na?ve T cells (CD4+CD45RBhigh) purified from spleens of untreated mice as described in the Supporting information. Levels of IL-10, IL-12p40 and interferon (IFN)-�� were measured in the culture supernatants using Bio-Plex cytokine assay kits (Bio-Rad). Data are expressed as mean?��?SD. The Instat package from GraphPad Software (San Diego, CA, USA) was used to analyse the data using the t-test with Welch's correction. We investigated the effect of EFD BCG on atherogenesis in Ldlr?/? and Apoe?/? mice. Independent of age, mouse strain and number of injections, EFD BCG treatment reduced lesion cross-sectional area in the aortic roots and decreased lipid lesion area in the en face aortic preparation compared http://www.selleck.cn/products/Bortezomib.html with treatment with PBS (Table?1 and Fig.?1). Immunohistochemical staining showed less infiltration of MOMA2?+?macrophages in atherosclerotic lesions in EFD BCG- compared with PBS-treated Ldlr?/? (Fig.?2a, b) and Apoe?/? mice (77.8?��?5.6 vs. 42.6 ��?7.4; P? http://www.selleckchem.com/products/PD-0332991.html cytokines [13]. Unphosphorylated retinoid X receptor �� (RXR��) forms a heterodimer with PPAR-�� to reduce inflammation [14]. EFD BCG increased PPAR-�� binding activity in vascular tissues and spleens and inhibited RXR�� phosphorylation in spleens of Ldlr?/? mice (Fig.?3a�Cc). Moreover, splenic nuclear extracts from EFD BCG-treated Ldlr?/? and Apoe?/? mice had higher levels of nuclear SP-1 (Fig.?3d and Fig. S1B), which is an important component of IL-10-mediated immunoregulation [15]. We analysed the effect of EFD BCG on transcription factors that regulate T-cell commitment.