Figure 4

Nilotinib inhibits the cytokine production by CD4⁺CD25⁺ T cells and CD4⁺CD25^- T cells. Panel A: CD4⁺CD25⁺ T cells were stimulated with anti-CD3, anti-CD28 and IL-2 in the presence or absence of 25 μM nilotinib. After 4 days incubation, supernatants were collected for cytokine analysis. The high intensity spots in the three corners are positive controls, the lower right contains the negative controls. Panel B: Graphs of the relative density (subtracted the average background signals) are shown for selected cytokines. Array images are shown in panel A and profiles created by quantifying the background-subtracted mean spot pixel densities are identified as shown in Panel B by using image analysis software. Panel C + D: The cytokine production of CD4⁺CD25^- T cells treated with nilotinib was analyzed by above methods.