The effect of Gli2ΔN2 expression and Gli2ΔC2 expression on Rag1−/− FTOC stimulated with anti-CD3. (A) Right bar chart shows thymocyte number in E15.5 GliΔN2Rag1−/− FTOC (n = 5) and littermate Rag1−/− FTOC (n = 4), after treatment for 3 days with 1 μg/mL anti-CD3 mab. The difference was statistically significant (P = .008). Dot plots show CD4 and CD8 expression on Gli2ΔN2Rag1−/− FTOC (n = 5) and littermate Rag1−/− FTOC (n = 4), after treatment for 3 days with 1 μg/mL anti-CD3 mab. Left bar chart shows the relative proportion of DP cells in the same experiments. The difference between GliΔN2Rag1−/− and littermate Rag1−/− is significant (P = .009). Error bars show SD. (B) ERK phosphorylation in anti-CD3–treated Rag1−/− and GliΔN2Rag1−/− thymocytes. Histograms of icphospho-ERK levels in control (untreated) and anti-CD3–treated (5 × 106cells/mL activated with 10 μg/mL anti-CD3 mab cross-linked with biotinylated anti–hamster IgG for 2 minutes) thymocytes from Rag1−/− and GliΔN2Rag1−/−. Bar chart shows mean fluorescence intensity (MFI) of icphospho-ERK staining in control and anti-CD3–stimulated Rag1−/− (n = 3) and GliΔN2Rag1−/− (n = 3) thymocytes. (C) Dot plots show CD4 and CD8 expression on E15.5 Rag1−/− (n = 10) and GliΔ2C2Rag1−/− (n = 10) FTOC after treatment for 3 days with 1 μg/mL anti-CD3 mab. Bar chart shows the relative proportion of DP cells for anti-CD3–treated Gli2ΔC2Rag1−/− FTOC and littermate Rag1−/− FTOC. The difference is significant (P = .001). Error bars show SD. (D) Relative thymocyte number in Gli2ΔC2Rag1−/− FTOC and littermate Rag1−/− FTOC, 3 days after anti-CD3 treatment. There was no significant difference. Error bars show SD. (E) Relative change in ERK phosphorylation in anti-CD3–treated Gli2ΔC2Rag1−/− (5 × 106cells/mL activated with 10 μg/mL anti-CD3 mab cross-linked with biotinylated anti–hamster IgG for 2 minutes; and Gli2ΔN2Rag1−/−, n = 3) thymocytes, normalized to that of their Rag1−/− littermate controls. Error bars show SE.
