Fig. 5.

Lack of 22(R)-OHC-stimulated-cholesterol efflux in S1P3-deficient macrophages. A: ³H-cholesterol efflux to ApoA-I was measured in S1P3 KO and WT macrophages incubated with 6.25 μM 22(R)-OHC in the presence and absence of 5 μM SKi for 18 h. Individual points represent individual mice. Data are presented as mean ± SEM (n = 3–10 per group). Statistical analysis was performed using two-way ANOVA followed by Bonferroni test. *** P < 0.001. B: Abca1 gene expression was analyzed by real-time PCR in S1P3 KO and WT macrophages treated with 6.25 μM 22(R)-OHC in the presence and absence of 5 μM SKi for 18 h. Individual points represent individual mice. Data are presented as mean ± SEM (n = 10–15 per group). Statistical analysis was performed using two-way ANOVA followed by Bonferroni test. * P < 0.05; *** P < 0.001. C: ABCA1 protein expression was analyzed by Western blotting in S1P3 KO and WT macrophages treated as in B. Quantification was performed in relation to vinculin. Individual points represent individual mice. Data are presented as mean ± SEM (n = 5 or 6 per group). Statistical analysis was performed using two-way ANOVA followed by Bonferroni test. * P < 0.05; *** P < 0.001. D: ³H-cholesterol efflux to ApoA-I was measured in C57Bl6 macrophages incubated with 6.25 μM 22(R)-OHC in the presence and absence of 1 μM S1P for 18 h. Individual points represent individual mice. Data are presented as mean ± SEM (n = 5–7 per group). Statistical analysis was performed using one-way ANOVA followed by a Student-Newman-Keuls test. * P < 0.05. n.s., not significant.