Figure 5. RORγt Inhibits Activation-Induced Fas Ligand Upregulation in KMIs-8.3.5 Cells.
(A) FACS analysis of cell surface expression of Fas, Fas ligand, and CD69 in KMIs-8.3.5hCD2 control or KMIs-8.3.5RORγt cells after TCR-mediated activation. KMI-8.3.5 cells expressing RORγt and control cells were activated on 2C11-coated plates for 4.5 hr and the cell surface phenotype was examined by FACS (filled, before stimulation; unfilled, after stimulation). A similar pattern was observed when PMA (10 ng/ml) and ionomycin (0.2 μg/ml) were used as stimuli (data not shown).
(B) Northern blot analysis of RNA expression of Fas ligand, Nur-77, and Egr-3 in KMIs-8.3.5hCD2 control, or KMIs-8.3.5RORγt cells after TCR-mediated activation. Cells were activated in 2C11-coated plates (125 ng/ml) for the indicated time, harvested, and extracted for total RNA. 15 μg total RNA was used for each sample. GAPDH serves as a loading control.