Figure 2
Figure 2. Activation of T cells through NKG2D and CD3 activates β-catenin. chNKG2D (ch) and wtNKG2D (wt) T cells were stimulated with media, anti-CD3/CD28–, anti-CD3–, anti-NKG2D–coated beads, or beads coated with both anti-CD3 and anti-NKG2D Abs. (A) Phosphorylated (Ser9) GSK3β expression in chNKG2D or wtNKG2D T cells was measured 30 minutes after stimulation. Data shown were obtained from 1 donor and are representative of data from 3 to 5 donors. (B) β-Catenin was measured with anti–β-catenin Abs at the indicated time points. Data shown were obtained from 1 donor and are representative of data from ≥ 3 donors. (D) Gene expression by chNKG2D and wtNKG2D T cells was determined by RT-PCR 8 hours after stimulation. Data are shown as the fold change in gene expression compared with T cells cultured in media. Stimulation of chNKG2D T cells through NKG2D significantly changed gene expression compared with CD3 stimulation (**P < .05), and stimulation through CD3 and NKG2D significantly changed gene expression compared with CD3 stimulation alone (***P < .05). Data shown were obtained from 1 donor and are representative of data from ≥ 4 donors. (D) ChNKG2D and wtNKG2D T cells were stimulated, and CD44 cell surface expression was determined by FACS after 24 or 72 hours. Data shown were obtained from 1 donor and are representative of data from ≥ 3 donors.

Activation of T cells through NKG2D and CD3 activates β-catenin. chNKG2D (ch) and wtNKG2D (wt) T cells were stimulated with media, anti-CD3/CD28–, anti-CD3–, anti-NKG2D–coated beads, or beads coated with both anti-CD3 and anti-NKG2D Abs. (A) Phosphorylated (Ser9) GSK3β expression in chNKG2D or wtNKG2D T cells was measured 30 minutes after stimulation. Data shown were obtained from 1 donor and are representative of data from 3 to 5 donors. (B) β-Catenin was measured with anti–β-catenin Abs at the indicated time points. Data shown were obtained from 1 donor and are representative of data from ≥ 3 donors. (D) Gene expression by chNKG2D and wtNKG2D T cells was determined by RT-PCR 8 hours after stimulation. Data are shown as the fold change in gene expression compared with T cells cultured in media. Stimulation of chNKG2D T cells through NKG2D significantly changed gene expression compared with CD3 stimulation (**P < .05), and stimulation through CD3 and NKG2D significantly changed gene expression compared with CD3 stimulation alone (***P < .05). Data shown were obtained from 1 donor and are representative of data from ≥ 4 donors. (D) ChNKG2D and wtNKG2D T cells were stimulated, and CD44 cell surface expression was determined by FACS after 24 or 72 hours. Data shown were obtained from 1 donor and are representative of data from ≥ 3 donors.

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