Fig. 3. SC synergistically reduced LPS and IFN-γ-induced NO production in murine RAW 264.7 cells (n≥3 individual experiments). (A) Dose-response curves of S, C and SC in inhibiting NO and (B) CI-Fa curve in RAW 264.7 cells. The CI-Fa curves revealed the interaction (synergistic when CI<1 or antagonistic when CI>1) of SC at all NO inhibitory levels (Fa values ranged from 0 to 1). (C) S, C and SC at 2.5 µg/mL inhibited LPS activated iNOS expression in the RAW 264.7 cells by immunofluorescence staining. Images were taken by a confocal microscope with 40x magnification. Blue: DAPI in the nucleus, green: iNOS in the RAW 264.7 cells. Scale bar=20 µm. (D) Statistical analysis of iNOS positive and negative fluorescence (% of total cells per image). #p<0.05 LPS vs. blank. ****p<0.0001 of iNOS positive compared with LPS. (E) Western blot analysis of iNOS total protein expression. RAW 264.7 cells were treated with medium only (Blank), LPS (1 µg/mL), S (5 µg/mL), C (5 µg/mL), and SC (5 µg/mL) for 24 h, and the total protein was lysed for Western blotting. #p<0.05 vs. Blank, *p<0.05 vs. LPS.
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