Figure 7.

Knockdown of ASM blocks CD3/CD28 signals and Th responses. (a–d) Control knockdown (sh-C) and ASM knockdown (sh-1 and sh-2) of healthy blood CD4⁺ T cells were stimulated with anti-CD3/CD28 antibody-coated beads, followed by representative TLC analyses of ceramide at 120 min (a), CD3/CD28 signaling at 120 min by western blotting (b), and FACS (fluorescence-activated cell sorting ) analysis (c), and CFSE expression at 96 h (d). (e) ASM knockdown of CD4⁺ T cells above were cultured under Th1-, Th2-, and Th17-polarizing conditions, and the intracellular levels of IFNγ, IL-4, and IL-17 were determined by FACS at 96 h, respectively. Data are representative of three independent experiments