Figure 5

Effects of n3 and n6 PUFA and COX-2 inhibition on FAS mRNA expression. 3T3 L-1 adipocytes were grown 6–7 days post-confluence, and starved for 24 h in serum-free media containing FA-free BSA. Fatty acids were incubated in media containing FA-free BSA for two hours at 37°C in a shaking water bath prior to treatment. Treatment consisted of CI (1 μM), EPA and AA (150 μM each treatment), and AA + EPA (75 μM each FA). After 48 hours, cells were scraped using 350 μL of Qiazol lysis reagent, and total RNA was extracted using the RNeasy™ lipid tissue midi kit (Qiagen) following the manufacturer's protocol. RNA was stored at -80° for further analysis. Real time RT-PCR analysis was performed as described in the materials and methods. For all treatments n = 6. Results represent the mean ± SEM. All values were significantly different than control, and values labeled with different letters are significantly different (p < 0.05) from each other. Values with the same letters do not differ significantly.