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Figure S1. Alignment of α2-helix forming amino acids of published CD1d sequences, showing conservation of W155 in catarrhines (apes and Old World monkeys) and of G153 in nearly all other species. 1. Pseudogene. 2. CD1 protein, but not orthologous to any specific eutherian isoform.

Figure S2. Gating strategy for enumeration of iNKT populations in lipid-treated PBMC cultures (as used for generating data shown in figure 1a). Shown is an example of a KRN7000 stimulated in vitro culture: Forward/side scatter characteristics were used to define region 1 (R1) containing lymphocytes (left density plot). Propidium iodide exclusion and staining with FITC-conjugated anti-CD3 antibody were used to define region 2 (R2), and from this live T lymphocytes were identified by gating cells on (R1+R2)(middle density plot), and live iNKT cells were identified as αGC-CD1d tetramer positive cells in gate(R1+R2) (right density plot). All flow cytometry data were acquired on a BD FACSCalibur, and data were analysed using CellQuest Pro software.

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