Antitumor effect of supernatants from C. crepidioides -stimulated RAW264.7 cells on S-180 cells and effects of C. crepidioides on NO production in RAW264.7 cells. (A, B) Antitumor effects of supernatants from C. crepidioides-stimulated RAW264.7 cells against S-180 cells, measured by WST-8 assay. RAW264.7 cells were cultured with various concentrations of C. crepidioides extract for 72 h. The control samples did not include RAW264.7 cells. At the end of each incubation period, the supernatant was collected. S-180 cells were precultured for 24 h. Thereafter, the culture media were removed and replaced with supernatants from C. crepidioides-stimulated RAW264.7 cells (A) or by the control medium (B). S-180 cells were incubated for another 24, 48 or 72 h, and cell growth was determined in triplicate cultures by WST-8 assay. Cell growth in control cultures (supernatants from unstimulated RAW264.7 cells (A) or control medium in the absence of C. crepidioides (B)) was considered 100%. Data are mean ± SD values expressed as percentage of the control. (C, D) Effects of C. crepidioides on NO production in RAW264.7 cells. RAW264.7 cells were incubated with or without C. crepidioides (500 μg/ml) (C) or with various concentrations of C. crepidioides (D) for the indicated time periods. NO production was determined by measuring the accumulation of nitrite in the culture medium. Data are mean ± SD of triplicate cultures.