Table 2.
Differentiation of RAW 264 7 cells toward secreting-activated macrophages following culture with LPS, IFN-γ or LPS+IFN-γ
| nitric oxide metabolism | ||||||||||||
| TNF production (pg/ml) | nitrites/nitrates (μM) | citruline (μM) | ||||||||||
| Sample type | 0 h | 6 h | 24 h | 48 h | 0 h | 6 h | 24 h | 48 h | 0 h | 6 h | 24 h | 48 h |
| medium | 156+/ 12 | 215+/ 18 | 332+/ 168 | 462 +/ 250 | ND | ND | ND | ND | 20+/ 4 | 22+/ 5 | 25+/ 7 | 33+/ 8 |
| LPS | 156+/ 12 | 1480+/ 187 | 1646+/ 266 | 1733+/ 284 | ND | ND | 26+/ 5 | 38+/ 18 | 20+/ 4 | 35+/ 12 | 47+/ 11 | 86+/ 31 |
| IFN | 156+/ 12 | 402+/ 28 | 681 +/ 42 | 819+/ 126 | ND | ND | 4+/ 2 | 12+/ 3 | 20+/ 4 | 25+/ 5 | 34+/ 9 | 42+/ 8 |
| LPS/IFN | 156+/ 12 | 1392+/ 224 | 1502+/ 335 | 1754+/ 201 | ND | ND | 13+/ 9 | 33+/ 15 | 20+/ 4 | 37+/ 12 | 43+/ 7 | 72+/ 22 |
ND: not detectable RAW 264.7 cells (250 000 cells/well) were cultured with medium or medium supplemented with either IFN-γ (50 U/ml), or LPS (5 μg/ml) or a mixture of IFN-γ (50 U/ml) + LPS (5 μg/ml). Cell-free culture supernatants were harvested at 0, 6, 24, and 48 hours. The concentrations of TNF-α (measured using an ELISA kit), nitrites/nitrates (measured using the Griess reagent) and citrulline (measured using colorimetric reaction), were evaluated in each culture supernatant as described in Methods. Data represent means +/- SEM of five independent experiments.