FIGURE 7.
Enhanced atherosclerosis development in PLA2G3 Tg × apoE-/- mice following a high fat diet. A, PLA2G3 Tg or LNL-PLA2G3 Tg mice were crossed with apoE knock-out (apoE-KO) mice. Blood samples were collected from the tail veins of PLA2G3tg/0 × apoE-/- (III-Tg), LNL-PLA2G3tg/0 × apoE-/- (LNL), and PLA2G3non-Tg × apoE-/- (-) mice as well as WT C57BL/6 (apoE+/+) mice, and plasma cholesterol concentrations were measured. B, PLA2 enzyme activity in the plasma was measured with 1-palmitoyl-2-[14C]linoleoylphosphatidylethanolamine as substrate. C, female mice were fed with a high fat/high cholesterol diet for 10 weeks, and plasma and aortas were collected. Concentrations of LPC in plasma (left) and in LDL (right) were measured using an enzyme-based kit. In A–C, blood samples for individual analyses were randomly chosen, which gave reasonable results in the first round of assays. D, aliquots of LDL fractions (10 μg of protein equivalent) along with size markers (ferritin, 12.2 nm; thyroglobulin, 17 nm; latex beads, 30 nm) were subjected to native PAGE. The gels were stained with Coomassie Brilliant Blue. Aggl, aggregated LDL. E and F, the whole aortas obtained from male and female mice fed a high fat diet for 10–14 weeks were stained with oil red O to visualize atherosclerotic lesions. The percentage of surface lesion area was calculated using Canvas software. Photographs of a typical example of each group are shown in E, and the percentage of surface lesion area in individual mice tested so far as well as the mean ± S.D. in each group is indicated in F. Data were statistically evaluated using one-factor analysis of variance and the Tukey-Kramer or Kruskal-Wallis test. Data with p < 0.05% were regarded as significant.