Figure 4. Protection of T Hybridomas from Activation-Induced Apopotosis by RORγt Deletion Mutants or RORγ.
(A) Both the DBD and the LBD of RORγt are required to protect T cell hybridomas from activation-induced cell death. DO11.10 or KMIs-8.3.5 cells expressing full-length or truncated RORγt or hCD2 were cultured in 2C11-coated plates (125 ng/ml) and tested for their [3H]Thymidine incorporation. Incorporation of cells cultured in medium alone is calculated as 100%. Numbers indicate the amino acids contained in each RORγt deletion mutant. An initiation methionine was added to RORγt deletion mutant 113–495.
(B) FACS analysis of hCD2 expression on KMIs-8.3.5 cells expressing RORγ or RORγt. Polyclonal KMIs-8.3.5 cell lines expressing RORγ or RORγt were generated by panning on anti-hCD2 MAb-coated plates, and adherent cells were stained with a FITC-anti-hCD2 as indicated. The profile at the far left represents parental KMIs-8.3.5 cells stained with anti-hCD2 MAb.
(C) Comparison of the anti-apopototic effects by RORγ and RORγt. KMIs-8.3.5 cells expressing RORγ (diamonds), RORγt (circles), or control cells (squares) shown in (B) were cultured in 2C11-coated plates and tested as described in (A).