Figure 2.
Mouse LIGHT is rapidly upregulated on activated T cells after polyclonal stimulation. (A) Naïve C57BL/6 splenocytes (2 × 105) were left untreated or were stimulated with PMA (100 ng/ml) and ionomycin (500 ng/ml) for 5 h. 1 μg/well Hilyte-647-labeled anti mouse LIGHT (3D11) or Hylite-647-labeled isotype control mouse IgG2b were added to cultures during incubation in a final volume of 250 μl. A lineage cocktail to gate out CD19+ CD11c+ Ly6G+ cells was included in the staining. The expression of mouse LIGHT was then analyzed on live resting and activated CD4 and CD8 T cells. (B) The expression of mouse LIGHT on the surface of resting or PMA plus ionomycin activated CD4+ and CD8+ T cells was also determined with the anti-LIGHT antibody pre-incubated with Flag-Foldon LIGHT (FF-LIGHT) (2 μg FF-LIGHT per well). The percentage of each population is represented in each quadrant. PMA: Phorbol Myristate Acetate. Iono: Ionomycin.