Figure 5: GHcer dampened the upregulation of CD25 and Notch1 during lymphocyte activation. Mouse splenocytes (A and C) and purified CD4+ T cells (B and D) were incubated with GHCer, ceramide or medium control for 24 hrs, and then activated by anti-mouse CD3 and CD28 mAb. Two days after activation, CD4+ T cells were gated to determine the expression of CD25 (A and B) and Notch1 (B and D) by flow cytometry. Purified mouse splenic CD19+ B cells were incubated with GHCer, ceramide or medium control for 24 hrs, and then activated by LPS combined with IL-4 and mouse CD40 ligand. Three days after activation, Notch1 expression was determined by flow cytometry (E). Human PBMC were incubate with GHCer or Cer for 24 hrs, and then activated by anti-human CD3 and CD28 mAb. Two days after activation, Notch1 expression on CD4+ T cells was determined by flow cytometry (F).