Fig 2: Expression of ApoM in Ana‐1 cells, peritoneal macrophages, liver, and kidney from WT mice. (A) The expression level of ApoM mRNA in Ana‐1 cells, peritoneal macrophages, liver, and kidney from WT mice. Data were normalized to the levels of ApoM in Ana‐1 cells as 100%. (***P < 0.01). (B) Capillary western immunoassay of ApoM protein in Ana‐1 cells, peritoneal macrophages, liver, and kidney from WT mice. Data were normalized to the levels of ApoM in Ana‐1 cells as 100%. (*P < 0.5). (C) Agarose gel electrophoresis of ApoM PCR product derived from Ana‐1 cells, peritoneal macrophages, liver, and kidney from WT mice. The middle lane contains DNA size markers. Lanes kidney and liver were the positive controls. (D) The sequencing of the PCR product in macrophages from WT mice. The PCR products were sequenced forward. Both probe and reverse‐primer sequences were matched with the products. Values are mean ± SD (n = 6 per group). Statistical analysis was performed using one‐way ANOVA test.

Fig 3: Effects of rh‐ApoM on cholesterol efflux from Ana‐1 cells. Ana‐1 cells were incubated with 10 μm NBD‐cholesterol for 4 h. A concentration of 20 μg·mL−1 ApoM+HDL (ApoM−HDL) or a combination of both ApoM+HDL (ApoM−HDL) and rh‐ApoM was used to induce cholesterol efflux. A set of cells was incubated with serum‐free medium as a control. Values are mean ± SD (n = 6 per group). Statistical analysis was performed using one‐way ANOVA test. *P < 0.001 vs. ApoM−HDL. ns not significantly different from ApoM+HDL (P > 0.05). efflux (%) = fluorescence intensities of supernatant/(fluorescence intensities of supernatant + fluorescence intensities of the cell lysate) × 100%.

Fig 4: Cholesterol efflux from Ana‐1 cells with different ApoM‐inducing solutions. Ana‐1 cells were incubated for 4 h in serum‐free medium supplemented with 10 μm NBD‐cholesterol. (A) The labeled cells were incubated with 20 μg·mL−1 ApoM+HDL, 20 μg·mL−1 ApoM−HDL, or 10 μg·mL−1 rh‐ApoM for another 0–8 h. (B) The labeled cells were incubated with 10–100 μg·mL−1 rh‐ApoM for 4 h. A set of cells was incubated with serum‐free medium as a control. Values are presented as the mean ± SD (n = 3–6 per group). Statistical analysis was performed using two‐way ANOVA followed by Bonferroni test and one‐way ANOVA test. *Significantly different from the control (P < 0.05), § Significantly different from ApoM−HDL (P < 0.05). ns not significantly different from the control (P > 0.05). efflux (%) = fluorescence intensities of supernatant / (fluorescence intensities of supernatant + fluorescence intensities of the cell lysate) × 100%.