Fig. 1.

Ligands for NKG2D are up-regulated in spontaneously arising lymphomas. (A) Splenocytes from Eμ-Myc mice with lymphoma and littermate controls (LMC) were stained with B220 (B cell marker) and mNKG2D/Fc and analyzed by flow cytometry. The histograms shown, if representing gated populations, are as indicated (e.g., B220+). The shaded histogram represents isotype control for mNKG2D/Fc, and the heavy black line represents mNKG2D/Fc staining. Log fluorescence is plotted on the x axis, and the percentage of maximum events per cells is plotted on the y axis. This scaling is maintained for all subsequent histograms. (B) (Left) Splenocytes from Eμ-Myc mice with lymphoma were stained with B220 and mNKG2D/Fc. The dotted line is mNKG2D/Fc staining after blocking with anti-mouse Rae-1ε, the heavy black line is mNKG2D/Fc staining after blocking with isotype control for anti-mouse Rae-1ε, the thin black line is mNKG2D/Fc staining alone, and the shaded histogram is isotype control for mNKG2D/Fc. (Center) Splenocytes from Eμ-Myc mice with lymphoma were stained with mNKG2D/Fc (heavy black line)/isotype control (shaded histogram) or (Right) anti-mouse Rae-1ε (heavy black line). The shaded histogram here represents anti-mouse Rae-1α/β/γ, and the thin black line represents an additional isotype control. Data are representative of three separate experiments. (C) Quantitative PCR analysis of Raet1 mRNA induction in B cells from wild-type (WT), Eμ-Myc preneoplastic, and Eμ-Myc tumor cells. Values are normalized to Ubiquitin gene expression and reflect fold induction over wild-type B cells (assigned a value of 1.0 and depicted in column 1).