Figure 9. Smad4 binds to the Eomes promoter to repress transcription.

• A–C. C57BL/6 CD8⁺ cells stimulated with anti-CD3/CD28 for 3 days were transfected with the Eomes luciferase reporter construct with various dosages of Smad4, Smad2, Smad3 and Smad4 or the various truncated Eomes luciferase reporter constructs with or without Smad4. TGF-β1 was treated as a control.
• D. CD8⁺ cells from Cd4Cre;Smad4^+/+ and Cd4Cre;Smad4^fl/fl mice were stimulated with anti-CD3/CD28 for 3 days, lysed and immunoprecipitated with either anti-Smad4 or rabbit IgG. Bound DNA was measured by qPCR using primers specific to the Eomes promoter. Graphs show mean + SEM (n = 3). Differential occupancy fold changes from four independent experiments are shown.