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Fig. 7. TCR down-regulation induced by strong and weak agonists. Fourteen hours after i.p. injection with 2 m g of the indicated antigens, livers were harvested and cell suspensions were prepared. Cells were labeled with CD1d/a -GalCer tetramer and mAb specific for a b TCR, NK1.1, and CD69. Density plots show CD1d/a -GalCer tetramer versus a b TCR expression by lymphoid-gated cells. The number represents the mean proportion of lymphoid-gated cells within the region indicated. Each column represents a sample from a group of mice treated with vehicle or glycolipid as indicated. Numbers represent the mean proportion ± standard deviation from three mice in each group. These data are representative of two separate experiments.

Fig. 8. Specific CD1d tetramer staining of liver mononuclear cells. Single-cell suspensions were prepared from the liver of 6- to 10-week-old C57BL/6J mice were stained for 2 h at room temperature, with glycolipid loaded or unloaded CD1d tetramers (final concentration 4 nM) and 10 m g/ml anti-TCRb mAb (PharMingen). Cells were then washed two times and analyzed by flow cytometry. One representative experiment of five is shown.