Fig. 3.

Effect of Cl-EE on iNOS in LPS-activated RAW264.7 cells. a Effect of Cl-EE on iNOS enzyme activity. RAW264.7 cells were pretreated with LPS (10 ng/mL) for 12 h and then exposed to Cl-EE at the indicated concentrations for another 12 h in the absence of LPS. The nitrite levels in the medium were measured by Griess method. b Effect of Cl-EE on iNOS mRNA expression. RAW264.7 cells were pretreated with the indicated concentrations of Cl-EE for 1 h and then exposed to LPS (10 ng/mL) for 4 h. The mRNA level of iNOS was determined by RT-PCR analysis. c Effect of Cl-EE on iNOS protein level. RAW264.7 cells were pretreated with Cl-EE at the indicated concentrations for 1 h and then exposed to LPS (10 ng/mL) for 24 h. Cellular proteins were extracted and the translation level of iNOS was determined by Western blot analysis. ^##P < 0.01 versus NC group; ^*P < 0.05 and ^**P < 0.01 versus LPS alone