(A) Expression of sLeX on CD8α+ T cells after transfections with control or CEBPD siRNA. CD8α+ T cells were divided into subsets as in Figure 1B and shown in Figure 2—figure supplement 1. Percentages of cells staining positive are indicated by the horizontal lines. Unless otherwise noted, experiments in this figure used the pool of four siRNAs, and in all experiments using siRNAs cells were harvested 3–4 days after transfections. (B) Expression of sLeX on T cells from multiple donors treated as in (A). (C) Relative expression of FUT7 and ST3GAL4 mRNAs in CD8α+ T cell subsets after transfections with control or CEBPD siRNAs. Each symbol type shows results from an individual donor. Red and green symbols are from experiments using non-overlapping pools of four or three CEBPD siRNAs, respectively, and blue symbols are from experiments using one of the siRNAs from the pool of three. (D) ChIP analysis of conventional memory-phenotype T cells and MAIT cells using anti-C/EBPδ antibodies and primers for amplifying sequences 5’ to the transcription start sites (TSS) for FUT7 and ST3GAL4 at 1 kb intervals. Data are expressed as percent of input DNA, and the dashed line indicates ‘background’ signal based on results for the intergenic region, IGX1A. (E) Expression of CCR6 on T cells after transfections with control or CEBPD siRNAs. Percentages of cells staining positive are indicated by the horizontal lines. (F) MFIs for CCR6 on T cells treated as in (E). (G) Relative expression of CCR6 mRNA in CD8α+ T cell subsets after transfections with control or CEBPD siRNAs as in (C). (H) ChIP analysis as in (D), except using primers for CCR6. (I) Relative expression of CCR2 and CCR5 mRNAs in CD8α+ T cell subsets after transfections with control or CEBPD siRNAs. (A, D, E and H) Data are from one representative of six (A), five (D), left panel), six (D), right panel), six (E), and two (H) donors. (B, C, F, G and I) Bars show means and SEMs, and data are from cells from a total of six (B, C, F, and G), four (I, left panel), or three (I, right panel) donors, each represented by a unique symbol. The p values were calculated using the ratio paired t test. (B, C, F, and G) ns, not significant; *, p<0.05; **, p<0.01; ***, p<0.001.