Modulation of NK1.1, CD94/NKG2, Ly49, NKG2D, and CD28 on α-GalCer– or OCH-activated liver iNKT cells. (A) Cell surface expression of the indicated molecules was analyzed on electronically gated α-GalCer/CD1d⁺ iNKT cells on the indicated days after intraperitoneal injection of α-GalCer or OCH. One day after α-GalCer or OCH injection, both cell surface and intracellular expression of the indicated molecules were analyzed in electronically gated intracellular Vβ^2/7/8+ iNKT cells. The analysis gates are indicated by the gray line in dot plot panels. Bold lines indicate the staining with the respective mAb, and the thin lines indicate the staining with isotype-matched control Ig. Similar results were obtained from 3 independent experiments. (B) Existence of a cell population expressing intracellular α-GalCer/CD1d–specific TCR 1 day after α-GalCer injection. Liver MNCs were intracellularly stained with α-Gal-Cer–loaded recombinant soluble dimeric mouse CD1d:Ig and PE-conjugated anti–mouse IgG1 mAb, or PE-conjugated anti–mouse IgG1 mAb together with FITC-conjugated anti-NK1.1 mAb, 1 day after α-GalCer injection. Quadrant gates were set by staining with FITC-conjugated isotype-matched control and PE-conjugated anti–mouse IgG1 mAb.