Fig. 4

Effects of EEDL on NO and iNOS levels in LPS-stimulated RAW 264.7 cells. a Nitrite production secreted in the culture medium was monitored by the Griess reagent system. Values are means ± SD (n = 6) from three independent experiments and significant difference compared with blank control or LPS treated alone, ^##P < 0.01 vs. blank control group, **P < 0.01 vs. LPS-treated group. b RAW 264.7 cells were pretreated with EEDL (200 μg/mL) and 0.2 μg/mL LPS for 18 h. The mRNA expression of iNOS was detected by real-time RT-PCR. Values are means ± SD (n = 3) and significance compared with blank control or LPS treated alone, ^##P < 0.01 vs. blank control group, *P < 0.05, **P < 0.01 vs. LPS-treated group. c Intracellular NO was detected by confocal laser scanning microscope: (a) blank control; (b) cells pretreated with LPS; (c) cells pretreated with EEDL in the present of LPS; (d) cells pretreated with 100 μM L-NAME in the present of LPS. d The protein expression of iNOS was measured with Western blot. The optical density was captured by the Image-Pro Plus version 6.0. Values are means ± SD (n = 3) and significant difference compared with blank control or LPS treated alone, ^##P < 0.01 vs. blank control group, **P < 0.01 vs. LPS-treated group