Fig. 6.

CerS activity using the optimized assay procedure. A: Assays were performed using different amounts of protein from HEK cells overexpressing the indicated CerS; the acyl-CoA was chosen according to the specificity of the particular CerS (i.e., C24:1-CoA for CerS2, C20-CoA for CerS4, and C16-CoA for CerS5 and CerS6). CerS2 and CerS4 were assayed for 30 min, and CerS5 and CerS6 for 10 min. Results are means ± SEM; n = 2–5. B: Assays were performed using different reaction times. CerS2 and CerS4 were assayed with 40 μg of protein, and CerS5 and CerS6 with 10 µg of protein. Results are means ± SEM; n = 2–5.