Skip to main content
. Author manuscript; available in PMC: 2014 Jul 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2013 May 2;33(7):1513–1520. doi: 10.1161/ATVBAHA.113.301498

Figure 3. The exogenous supplementation of ceramide in culture significantly reducesPPARγ2 protein levels.

Figure 3

Huh7 cells were incubated with exogenous 0, 10, 30 μM of sphingomyelin, ceramide or phosphatidylcholine for 24 h. Cellular PPARγ2, CD36 and FSP27 mRNA levels were measured by real-time PCR. Panel A, PPARγ2 mRNA levels. Panel B, CD36 mRNA levels. Panel C, FSP27 mRNA levels. Cellular PPARγ2 mass were measured by Western blotting. Panel D and E, PPARγ2 Western blot after treatment with exogenous 0, 10, 30 μM sphingomyelin, ceramide or phosphatidylcholine for 24 h. Panel F and G, PPARγ2 Western blot after treatment with exogenous 0, 10, 30 μM of ceramides C16:0, C22:0, and C24:0 for 24 h. Panel H and I, PPAR reporter assay. Huh7 cells were transfected with PPAR reporter, negative control or positive control, respectively. After 24 h of transfection, the cells were treated with exogenous sphingomyelin, ceramide or phosphatidylcholine for 24 h, and then dual-luciferase reporter assay was performed. Panel H, dual-luciferase reporter assay after treatment with exogenous 0, 20, 40 μM of sphingomyelin, ceramide or phosphatidylcholine for 24 h. Panel I, dual-luciferase reporter assay after treatment with exogenous 0, 20, 40 μM of ceramides C16:0 and C24:0 for 24 h. Values are Mean ± SD., n=5, *P<0.05, **P<0.01.