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sji12039-sup-0001-FigureS1.pdfapplication/PDF22KFigure S1 DexVD3 DC have an anti-inflammatory cytokine profile. The cell-free supernatants were collected when harvesting the DC populations and stored in aliquots at −20 °C until analysis. The amount of cytokines/chemokines secreted were analyzed using a 25-plex bead assay and the concentrations are given in pg/ml. Cytokines and chemokines that showed results below the detection limit (IL-1β, IP-10, MCP-1, MIP-1a, MIP-1b) are not shown. GM-CSF and IL-4 were part of the added cytokines and therefore not considered in the analysis. Data are shown as mean ± SD and represent three independent experiments. See Fig. 1 for definitions.
sji12039-sup-0002-FigureS2.pdfapplication/PDF132KFigure S2 Gating strategy for the analysis of the suppressive capacity of T cells primed with various tolDC populations. Naïve responder cells were labelled with CFSE and tolDC-primed NAC were labelled with CellTrace Violet. This allowed exclusive gating on CellTrace Violet negative cells and further analysis of proliferation of CFSE-labelled responder cells. The top left dot plot depicts cells in a live gate based on forward scatter and side scatter, the top right histogram depicts gating on CellTrace Violet negative cells. The left bottom histogram shows proliferation of CFSE-labelled CellTrace Violet negative responder cells after co-culture with immature DMSO DC-primed NAC. The right bottom histogram is an overlay of proliferation histograms of CFSE-labelled CellTrace Violet negative responder cells after co-culture with different tolDC-primed NAC. MFI values for the different populations are shown in the legend.
sji12039-sup-0003-FigureS3.pdfapplication/PDF1939KFigure S3 Gating strategy for the analysis of NAC phenotype. The top row of dot plots shows cells in a live gate based on forward scatter and side scatter, the second row depicts gating on CD4+ cells based on unstained negative controls. 3rd row depicts the proportion of the CD25FoxP3 cells amongst the CD4+ NAC. 4th row shows the proportion of CD4+IL-10+CD25FoxP3- Tr1 cells. The bottom row shows gating on CD19+IL-10+ double positive Breg cells based on unstained negative controls. The DC population that was used to stimulate NAC is shown on the top. See Fig. 1 for definitions.
sji12039-sup-0004-FigureS4.pdfapplication/PDF25KFigure S4 Cytokine and chemokine profiling of culture supernatants from NAC primed with various DC populations. The cell-free culture supernatants were collected after co-culture of DC and NAC and stored in aliquots at −20 °C until analysis. The amount of cytokines/chemokines secreted was analyzed using a 25-plex bead assay and the concentrations are given in pg/ml. Cytokines and chemokines that showed results below the detection limit (IL-4, IP-10, IL-1β, MCP-1, MIP-1b) are not shown. Data are shown as mean ± SD and represent three independent experiments. Only differences in IL-12 production are statistically significant (P < 0.05). The tolDC population that was used to stimulate NAC is shown on the x-axis. See Fig. 1 for definitions.
sji12039-sup-0005-FigureS5.pdfapplication/PDF34KFigure S5 Cytokine and chemokine profiling of culture supernatants from suppression experiments. The supernatants were collected before the FACS analysis of proliferation of the responder cells and stored in aliquots at −20 °C until analysis. The amount of cytokines/chemokines secreted were analyzed using a 25-plex bead assay and the concentrations are given in pg/ml. Cytokines and chemokines that showed results below the detection limit (IL-1β, IL-4, IL-17, MIP-1a, MIP-1b) are not shown. Data are shown as mean ± SD and represent three independent experiments. The DC population that was used to stimulate the effector cells is shown on the x-axis. See Fig. 1 for definitions. CD25 defines the depletion of CD25+ from the effector population.

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