Journal: Bioscience Reports
Article Title: Characterization of secretory sphingomyelinase activity, lipoprotein sphingolipid content and LDL aggregation in ldlr −/− mice fed on a high-fat diet
doi: 10.1042/BSR20120036
Figure Lengend Snippet: Mice of the indicated genotypes were placed on either a standard or modified diet for 10 weeks. S-SMase activity was measured using C 6 -NBD-SM as a substrate. ( A ) S-SMase activity in C57Bl6 mice measured by using 0.4 nmol of NBD-SM per sample. Results are shown as means±S.D. ( n =3) for each mouse. ( B–D ) Michaelis–Menten kinetics of the S-SMase activity in C57Bl6 and ldlr −/− mice on either standard or modified diet. The assays are carried out using serum (4–6 animals per assay) and the indicated substrate concentrations. Mean values of triplicates ±S.D. are shown. * P <0.05 and ** P <0.01 according to a Student's t test. SM/NBD-SM ratio in ( D ) was calculated by dividing the measured endogenous LDL-SM concentration (presented in ) and the amount of NBD-SM in each assay.
Article Snippet: C 6 -NBD-Cer {6-[ N -(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]hexanoylceramide} and C 6 -NBD-SM {6-[ N -(7-nitro-2,1,3benzoxadiazol-4-yl)amino]hexanoylsphingosylphosphocholine} were from Molecular Probes, C 17:0 ceramide ( N -heptadecanoyl- D - erythro -sphingosine), C 17:0 SM ( N -heptadecanoyl- D - erythro -sphingosylphosphocholine), N -acetyl-C 20 -sphinganine and bovine brain ceramide were purchased from Avanti Polar Lipids. bSMase was from Sigma–Aldrich.
Techniques: Modification, Activity Assay, Concentration Assay