Figure 2.

Identification of CerS2 as major ceramide synthase in RAW264.7 mouse macrophages. (A) The reaction catalyzed by ceramide synthase. (B) Ceramide synthase in vitro activity assays on membrane lysates from RAW264.7 mouse macrophages treated with fumonisin (5 μM, 30 min) or DMSO, showing robust inhibition of ceramide synthase activity following fumonisin treatment. As a control, denatured membrane proteomes were used. Data represent mean ± SEM for three biological replicates per group. (C) Western blot analysis confirming abundant expression of CerS2 in RAW264.7 membrane lysates. As a positive control, membrane lysates from HEK293T were used. A total of 40 μg of lysate was loaded for all samples in this analysis, and actin was used as a loading control. The Western blot analysis was performed on five biological replicates with reproducible results.