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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
CYTO_22203_sm_SuppFig1.pdf103KSupplemental Figure 1: Gating strategy for T cell surface phenotyping. Umbilical cord T cells were identified using sequential gating for FSC/SSC/ doublet-/LiveDead-/CD14-/CD235-/CD3+ cells. CD4+, CD8+ and ?d-T cells were identified with positive cells markers. CD4 T cells were further phenotyped using chemokine receptor expression.
CYTO_22203_sm_SuppFig2.pdf311KSupplemental Figure 2: Gating strategy for T cell functional phenotyping, adult control sample. Plots a-c are from a representative adult sample. T cells were identified using sequential gating for FSC/SSC/ doublet-/LiveDead-/CD14-/CD3+ cells (a). Cytokine positive gating was set based on fluorescence minus one controls and multiple iterations to determine the best fit. CD69+ staining was used as an internal control to isolate analysis to activated cells in order to exclude non-specific antibody staining (CD4-b, CD8-c).
CYTO_22203_sm_SuppFig3.pdf259KSupplemental Figure 3: Gating strategy for T cell functional phenotyping, representative umbilical cord sample. Plots a-c are from a representative umbilical cord sample. T cells were identified using sequential gating for FSC/SSC/ doublet-/LiveDead-/CD14-/CD3+ cells (a). Cytokine positive gating was set based on fluorescence minus one controls and multiple iterations to determine the best fit. CD69+ staining was used as an internal control to isolate analysis to activated cells in order to exclude non-specific antibody staining (CD4-b, CD8-c).
CYTO_22203_sm_SuppFig4.pdf70KSupplemental Figure 4: Cytokine frequencies for unstimulated CD4+ and CD8+ T cells measured by flow cytometry. Unstimulated conditions for both CD4 (a) and CD8 (b) T cells produced low frequencies of spontaneous activation.
CYTO_22203_sm_SuppTab1.pdf45KSupplemental Table 1: LSR configuration
CYTO_22203_sm_SuppTab2.pdf38KSupplemental Table 2: T cell phenotype panel
CYTO_22203_sm_SuppTab3.pdf44KSupplemental Table 3: Intracellular cytokine staining panel
MIFlowCyt-Item-Checklist.doc4794KSupporting Information: MIFlowCyt

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